Methods of Treatment by Administering an Anti-BAFFR Antibody Therapeutic Formulation

ABSTRACT

Anti-BAFFR antibodies are formulated as liquid formulation comprising a high concentration of the antibody active ingredient for delivery to a patient without high levels of antibody aggregation. The aqueous pharmaceutical composition may include one or more sugars, a buffering agent, a surfactant, and/or a free amino acid.

TECHNICAL FIELD

The present invention relates to a pharmaceutical formulation of anantibody against BAFFR (BAFF receptor), a process for the preparationthereof and uses of the formulation.

BACKGROUND

The BAFFR:BAFF pair is critically involved in the maturation oftransitional B-cells, for survival and activation of mature B-cells, andfor isotype class switching in response to T cell-dependent antigens.BAFF and its receptor BAFFR are also important for survival and growthof malignant B-cells. Further, BAFFR normally is not expressed on pre-Bcells, but was recently shown to be expressed on human ALL (B-lineageacute lymphoblastic leukemia) cells (Parameswaran, 2010, Cancer Res.70(11) 4346-4356). The removal of autoreactive B cells and the blockadeof inappropriate survival/activation mediated by excess BAFF levels inpatients suffering from autoimmune disorders or cancer represents awell-validated therapeutic goal. Thus, an anti-BAFFR antibody, inparticular an antibody capable of antibody-dependent cell-mediatedcytotoxicity (ADCC) and blockade of ligand binding to BAFFR may offer aneffective therapeutic agent in autoimmune diseases and B cell neoplasms.

Antibodies against BAFFR are known from e.g. WO 2010/007082 and includeantibodies which are characterized by comprising a V_(H) domain with theamino acid sequence of SEQ ID NO: 1 and a V_(L) domain with the aminoacid sequence of SEQ ID NO: 2. The antibody MOR6654 is one such antibody(IgG1 kappa). It has the heavy chain amino acid sequence of SEQ ID NO: 9and the light chain amino acid sequence of SEQ ID NO: 10. This antibodymay be expressed from SEQ ID NOs: 14 and 15, preferably in a host cellwhich lacks fucosyl-transferase, for example in a mammalian cell linewith an inactive FUT8(−/−) gene, to provide a functional non-fucosylatedanti-BAFFR antibody with enhanced ADCC. This antibody is referred tohereafter as MOR6654B. Alternative ways to produce non-fucosylatedantibodies are known in the art.

Therapeutic antibodies are typically formulated either in aqueous formready for parenteral administration or as lyophilisates forreconstitution with a suitable diluent prior to administration.

International Application PCT/EP2011/072248 discloses lyophilisates,which can be reconstituted to give a solution with a high concentrationof the antibody active ingredient and a low level of antibodyaggregation for delivery to a patient. High concentrations of antibodyare useful as they reduce the dosing volume, which must be delivered toa patient. Reduced dosing volumes minimize the time taken to deliver afixed dose to the patient.

Pharmaceutical compositions formulated to contain high concentration ofantibody may, however, have short shelf lives and the formulatedantibodies may loose biological activity resulting from chemical andphysical instabilities during the storage. Among those, aggregation,deamidation and oxidation are known to be the most common causes ofantibody degradation. Further, aggregation can potentially lead toincreased immune response in patients, leading to safety concerns. Thusantibody aggregation in pharmaceutical compositions must be minimized orprevented.

It is therefore an objective of the present invention to provide furtherand improved pharmaceutical compositions comprising anti-BAFFRantibodies, formulated such as to allow high concentration of anti-BAFFRantibodies with no or substantially no antibody aggregation.

It is another objective of the present invention to provide ananti-BAFFR antibody formulation suitable for subcutaneousadministration. The advantage of subcutaneous injection is that itallows the medical practitioner to perform it in a rather shortintervention with the patient. Moreover the patient can be trained toperform the subcutaneous injection by himself.

This objective is met by the pharmaceutical aqueous compositions of thepresent invention.

The aqueous compositions of the invention comprise high concentration ofanti-BAFFR antibodies, but no or essentially no aggregated antibodiesand are thus particularly suitable for subcutaneous administration.

In one embodiment, the present invention relates to aqueous compositionshaving a pH of 5.0-7.0 and comprising

-   (i) an anti-BAFFR antibody wherein the antibody has a concentration    of 18-165 mg/mL, and wherein said anti-BAFFR antibody includes heavy    chain CDR1, CDR2 and CDR3 of SEQ ID NOs 3, 4 and 5 respectively, and    light chain CDR1, CDR2 and CDR3 of SEQ ID NOs: 6, 7 and 8,-   (ii) a stabilizer,-   (iii) a buffering agent,-   (iv) a surfactant, and, optionally,-   (v) an amino acid.

In particular, the invention provides an aqueous composition having a pHof 5.5-6.5 and

-   -   comprising

-   (i) an anti-BAFFR antibody wherein the antibody has a concentration    of 18-165 mg/mL, and wherein said anti-BAFFR antibody includes heavy    chain CDR1, CDR2 and CDR3 of SEQ ID NOs 3, 4 and 5 respectively, and    light chain CDR1, CDR2 and CDR3 of SEQ ID NOs: 6, 7 and 8,

-   (ii) sucrose, trehalose or mannitol as a stabilizer,

-   (iii) histidine, citrate or succinate as a buffering agent,

-   (iv) polysorbate 20, poloxamer 188 or hydroxyproyl-b-cyclodextrin as    a surfactant, and, optionally,

-   (v) arginine as an amino acid.

In one embodiment, the aqueous composition of the invention as describedherein in the various embodiments comprises the anti-BAFFR antibody in aconcentration of between 20 mg/mL and 150 mg/mL, particularly of between80 mg/mL and 150 mg/mL, particularly of between 100 mg/mL and 150 mg/mL.

In one embodiment, the aqueous composition of the invention as describedherein in the various embodiments comprises a sugar as a stabilizingagent, particularly sucrose, mannitol or trehalose, in a concentrationof between 80 mM and 300 mM, particularly of between 120 mM and 270 mM,particularly of between 120 mM and 220 mM.

In one embodiment, the aqueous composition of the invention as describedherein in the various embodiments comprises a surfactant, particularlypolysorbate 20 or poloxamer 188, in a concentration of between 0.01% and−0.1%, particularly of between 0.02% and 0.06%.

In one embodiment, the aqueous composition of the invention as describedherein in the various embodiments comprises a buffering agent,particularly histidine, citrate or succinate, in a concentration ofbetween 5 mM and 50 mM, particularly in a concentration of between 15 mMand 25 mM, particularly of between 18 mM and 22 mM, particularly 20 mM.

In one embodiment, the aqueous composition of the invention as describedherein in the various embodiments comprises in addition an amino acid,particularly arginine or arginine-HCl, in a concentration of between 2mM and 80 mM.

In one embodiment, the aqueous composition of the invention as describedherein in the various embodiments comprises sucrose or trehalose in aconcentration of between 110 mM and 250 mM.

In a specific embodiment, the present invention provides an aqueouscomposition having a pH of 5.5-6.5 and comprising

-   (i) an anti-BAFFR antibody wherein the antibody has a concentration    of 18 mg/mL-165 mg/mL, and wherein said anti-BAFFR antibody includes    heavy chain CDR1, CDR2 and CDR3 of SEQ ID NOs 3, 4 and 5    respectively, and light chain CDR1, CDR2 and CDR3 of SEQ ID NOs: 6,    7 and 8,-   (ii) 80 mM-300 mM sucrose, trehalose or mannitol as a stabilizer,-   (iii) 5 mM-50 mM histidine, citrate or succinate as a buffering    agent,-   (iv) 0.01%-0.1% polysorbate 20 or poloxamer 188, or 1 mM-3 mM    hydroxyproyl-b-cyclodextrin as a surfactant, and, optionally,-   (v) 2 mM-80 mM arginine, particularly arginine-HCl.

In another specific embodiment, the present invention provides anaqueous composition having a pH of 5.5-6.5 comprising

-   (i) an anti-BAFFR antibody wherein the antibody has a concentration    of 20 mg/mL-150 mg/mL and wherein said anti-BAFFR antibody includes    heavy chain CDR1, CDR2 and CDR3 of SEQ ID NOs 3, 4 and 5    respectively, and light chain CDR1, CDR2 and CDR3 of SEQ ID NOs: 6,    7 and 8,-   (ii) 110 mM-250 mM sucrose or trehalose as a stabilizer,-   (iii) 15 mM-25 mM histidine, citrate or succinate as a buffering    agent,-   (iv) up to 0.02%-0.06% polysorbate 20 or poloxamer 188 or 2 mM-3 mM    hydroxyproyl-b-cyclodextrin as a surfactant, and, optionally,-   (v) 2 mM-80 mM arginine, particularly arginine-HCl.

In still another specific embodiment, the present invention provides anaqueous composition having a pH of 5.5-6.5 and comprising

-   (i) an anti-BAFFR antibody wherein the antibody has a concentration    of 20 mg/mL-150 mg/mL and wherein said anti-BAFFR antibody includes    heavy chain CDR1, CDR2 and CDR3 of SEQ ID NOs 3, 4 and 5    respectively, and light chain CDR1, CDR2 and CDR3 of SEQ ID NOs: 6,    7 and 8,-   (ii) 120 mM-220 mM sucrose or trehalose as a stabilizer,-   (iii) 18 mM-22 mM histidine, citrate or succinate as a buffering    agent,-   (iv) up to 0.02%-0.06% polysorbate 20 or poloxamer 188 or 2.5 mM    hydroxyproyl-b-cyclodextrin as a surfactant, and optionally-   (v) 2 mM-80 mM arginine, particularly arginine-HCl.

In another specific embodiment, the present invention provides anaqueous composition having a pH of 6.0 and comprising

-   (i) an anti-BAFFR antibody wherein the antibody has a concentration    of 150 mg/mL and wherein said anti-BAFFR antibody includes heavy    chain CDR1, CDR2 and CDR3 of SEQ ID NOs 3, 4 and 5 respectively, and    light chain CDR1, CDR2 and CDR3 of SEQ ID NOs: 6, 7 and 8,-   (ii) 220 mM sucrose as a stabilizer,-   (iii) 20 mM histidine as a buffering agent,-   (iv) 0.04% polysorbate 20 as a surfactant.

In another specific embodiment, the present invention provides anaqueous composition having a pH of 6.0 and comprising

-   (i) an anti-BAFFR antibody wherein the antibody has a concentration    of 150 mg/mL and wherein said anti-BAFFR antibody includes heavy    chain CDR1, CDR2 and CDR3 of SEQ ID NOs 3, 4 and 5 respectively, and    light chain CDR1, CDR2 and CDR3 of SEQ ID NOs: 6, 7 and 8,-   (ii) 220 mM trehalose as a stabilizer,-   (iii) 20 mM histidine as a buffering agent,-   (iv) 0.04% polysorbate 20 as a surfactant.

In another specific embodiment, the present invention provides anaqueous composition having a pH of 6.0 and comprising

-   (i) an anti-BAFFR antibody wherein the antibody has a concentration    of 150 mg/mL and wherein said anti-BAFFR antibody includes heavy    chain CDR1, CDR2 and CDR3 of SEQ ID NOs 3, 4 and 5 respectively, and    light chain CDR1, CDR2 and CDR3 of SEQ ID NOs: 6, 7 and 8,-   (ii) 120 mM sucrose or trehalose as a stabilizer,-   (iii) 20 mM histidine as a buffering agent,-   (iv) 0.04% polysorbate 20 as a surfactant, and-   (v) 50 mM arginine, particularly arginine-HCl.

In another specific embodiment, the present invention provides anaqueous composition having a pH of 6.0 and comprising

-   (i) an anti-BAFFR antibody wherein the antibody has a concentration    of 20 mg/mL and wherein said anti-BAFFR antibody includes heavy    chain CDR1, CDR2 and CDR3 of SEQ ID NOs 3, 4 and 5 respectively, and    light chain CDR1, CDR2 and CDR3 of SEQ ID NOs: 6, 7 and 8,-   (ii) 220 mM sucrose or trehalose as a stabilizer,-   (iii) 20 mM histidine as a buffering agent, and-   (iv) 0.04% polysorbate 20 as a surfactant.

In one embodiment, the invention relates to the aqueous composition ofthe invention as described herein in the various embodiments, whereinthe anti-BAFFR antibody comprises a V_(H) domain with amino acid SEQ IDNO: 1 and a V_(L) domain with amino acid SEQ ID NO: 2.

In another embodiment of the invention, the aqueous composition of theinvention as described herein in the various embodiments, wherein theanti-BAFFR antibody comprises a heavy chain region of SEQ ID NO: 9 and alight chain region of SEQ ID NO: 10.

In one embodiment, the anti-BAFFR antibody is a non-fucosylatedanti-BAFFR antibody.

The invention further provides a delivery device comprising the aqueouscomposition of the invention as described herein in the variousembodiments.

This delivery device may be provided in form of a pre-filled syringecomprising the aqueous composition of the invention as described hereinin the various embodiments.

In one embodiment, the invention relates to a method for delivering ananti-BAFFR antibody to a mammal, comprising the step of administering tosaid mammal an aqueous composition of the invention as described hereinin the various embodiments, particularly in form of a delivery devicesuch as a pre-filled syringe.

The present invention further provides the composition or the deliverydevice, or the pre-filled syringe of the invention as described hereinin the various embodiments, for use in treating a disease or disorderthat is mediated by BAFF receptor or that can be treated by killing ordepleting B cells.

In particular, the pharmaceutical composition or the delivery device, orthe pre-filled syringe of the invention as described herein in thevarious embodiments may be used in the treatment of autoimmune diseases,B cell neoplasms, such as lymphoma, leukemia or myeloma, rheumatoidarthritis, systemic lupus erythematosus, Sjögren's syndrome or Pemphigusvulgaris.

The invention is based, at least partly, on the properties of formulatedantibodies such as MOR6654 and MOR6654B, which retain remarkablestability and bioactive properties when formulated in a highconcentration as a liquid (aqueous) composition.

As used herein, an “aqueous” pharmaceutical composition is a compositionsuitable for pharmaceutical use, wherein the aqueous carrier isdistilled water. A composition suitable for pharmaceutical use may besterile, homogeneous and/or isotonic. Aqueous pharmaceuticalcompositions may be prepared either directly in an aqueous form, forexample in pre-filled syringe ready for use (the “liquid formulations”)or as lyophilisate to be reconstituted shortly before use.

As used herein, the term “aqueous pharmaceutical composition” refers tothe liquid formulation or reconstituted lyophilized formulation. Incertain embodiments, the aqueous pharmaceutical compositions of theinvention are suitable for parenteral administration to a human subject.In a specific embodiment, the aqueous pharmaceutical compositions of theinvention are suitable for subcutaneous administration.

As used herein, the phrase “parenteral administration” means mode ofadministration other than enteral and topical administration, usually byinjection, and includes, without limitation, intravenous, intramuscular,intraarterial, intrathecal, intracapsular, intraorbital, intracardiac,intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular,intraarticular, subcapsular, subarachnoid, intraspinal, epidural andintrastemal injection and infusion.

The use of antibodies as the active ingredient of pharmaceuticals is nowwidespread, including the products HERCEPTIN™ (trastuzumab), RITUXAN™(rituximab), SYNAGIS™ (palivizumab), etc. Techniques for purification oftherapeutic antibodies to a pharmaceutical grade are well known in theart.

The composition will usually be non-pyrogenic e.g. containing <1 EU(endotoxin unit, a standard measure) per dose, and preferably <0.1 EUper dose. The composition is preferably gluten-free.

In specific embodiments, the aqueous pharmaceutical compositions of theinvention exhibit low to undetectable levels of antibody aggregation ordegradation, with very little to no loss of the biological activitiesduring manufacture, preparation, transportation and long periods ofstorage, the concentration of the anti-BAFFR antibody being at leastabout 50 mg/mL, 100 mg/mL, 150 mg/mL, 200 mg/mL, 250 mg/mL, or 300mg/mL.

In one aspect, the invention relates to an aqueous pharmaceuticalcomposition with high concentration of anti-BAFFR antibodies.

It is known in the art that such high concentration aqueouspharmaceutical compositions can be diluted prior to injection, forexample, if lower antibody concentrations are required for specifictherapeutic interventions or when treating patients of lower body weightincluding children. Suitable concentrations can be 25 mg/mL or 10 mg/mL.Alternatively, the original formulation may be produced with such alower concentration.

The term “antibody” as referred to herein includes whole antibodies andany antigen binding fragment (i.e., “antigen-binding portion”) or singlechains thereof. A naturally occurring “antibody” is a glycoproteincomprising at least two heavy (H) chains and two light (L) chainsinter-connected by disulfide bonds. Each heavy chain is comprised of aheavy chain variable region (abbreviated herein as V_(H)) and a heavychain constant region. The heavy chain constant region is comprised ofthree or four domains, depending on the isotype, C_(H)1, C_(H)2, C_(H)3and C_(H)4. Each light chain is comprised of a light chain variableregion (abbreviated herein as V_(L)) and a light chain constant region.The light chain constant region is comprised of one domain, C_(L). TheV_(H) and V_(L) regions can be further subdivided into regions ofhypervariability, termed complementarity determining regions (CDR),interspersed with regions that are more conserved, termed frameworkregions (FR). Each V_(H) and V_(L) is composed of three CDRs and fourFRs arranged from amino-terminus to carboxy-terminus in the followingorder: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of theheavy and light chains contain a binding domain that interacts with anantigen. The constant regions of the antibodies may mediate the bindingof the immunoglobulin to host tissues or factors, including variouscells of the immune system (e.g., effector cells) and the firstcomponent (Clq) of the classical complement system.

The term “antigen-binding portion” of an antibody (or simply “antigenportion”), as used herein, refers to full length or one or morefragments of an antibody that retain the ability to specifically bind toan antigen (e.g., a portion of BAFFR). It has been shown that theantigen-binding function of an antibody can be performed by fragments ofa full-length antibody. Examples of binding fragments encompassed withinthe term “antigen-binding portion” of an antibody include a Fabfragment, a monovalent fragment consisting of the V_(L), V_(H), C_(L)and C_(H)1 domains; a F(ab)₂ fragment, a bivalent fragment comprisingtwo Fab fragments linked by a disulfide bridge at the hinge region; a Fdfragment consisting of the V_(H) and C_(H)1 domains; a Fv fragmentconsisting of the V_(L) and V_(H) domains of a single arm of anantibody; a dAb fragment (Ward et al., 1989 Nature 341:544-546), whichconsists of a V_(H) domain; and an isolated complementarity determiningregion (CDR).

Furthermore, although the two domains of the Fv fragment, V_(L) andV_(H), are coded for by separate genes, they can be joined, usingrecombinant methods, by a synthetic linker that enables them to be madeas a single protein chain in which the V_(L) and V_(H) regions pair toform monovalent molecules (known as single chain Fv (scFv); see e.g.,Bird et al., 1988 Science 242:423-426; and Huston et al., 1988 Proc.Natl. Acad. Sci. 85:5879-5883). Such single chain antibodies are alsointended to be encompassed within the term “antigen-binding region” ofan antibody. These antibody fragments are obtained using conventionaltechniques known to those of skill in the art, and the fragments arescreened for utility in the same manner as are intact antibodies.

An “isolated antibody”, as used herein, refers to an antibody that issubstantially free of other antibodies having different antigenicspecificities, e.g., an isolated antibody that specifically binds humanBAFFR is substantially free of antibodies that specifically bindantigens other than BAFFR. An isolated antibody that specifically bindsBAFFR may, however, have cross-reactivity to other antigens, such asBAFFR molecules from other species. Moreover, an isolated antibody maybe substantially free of other cellular material and/or chemicals.

The terms “monoclonal antibody” or “monoclonal antibody composition” asused herein refer to a preparation of antibody molecules of singlemolecular composition. A monoclonal antibody composition displays asingle binding specificity and affinity for a particular epitope.

The term “human antibody”, as used herein, includes antibodies havingvariable regions in which both the framework and CDR regions are derivedfrom sequences of human origin. Furthermore, if the antibody contains aconstant region, the constant region also is derived from such humansequences, e.g., human germline sequences, or mutated versions of humangermline sequences or antibody containing consensus framework sequencesderived from human framework sequences analysis, for example, asdescribed in Knappik, et al. (2000. J Mol Biol 296, 57-86).

The structures and locations of immunoglobulin variable domains, e.g.,CDRs, may be defined using well known numbering schemes, e.g., the Kabatnumbering scheme, the Chothia numbering scheme, a combination of Kabatand Chothia (AbM), etc. (see, e.g., Sequences of Proteins ofImmunological Interest, U.S. Department of Health and Human Services(1991), eds. Kabat et al.; Al Lazikani et al. (1997) J. Mol. Bio.273:927 948). Throughout this specification, the complementaritydetermining region (“CDR”) is defined according to the Kabat definitionwith the exception of CDRH1 which is the stretch of amino acids definedby a combination of both Kabat and Chothia definitions for this CDR.

The human antibodies of the invention may include amino acid residuesnot encoded by human sequences (e.g., mutations introduced by random orsite-specific mutagenesis in vitro or by somatic mutation in vivo).However, the term “human antibody”, as used herein, is not intended toinclude antibodies in which CDR sequences derived from the germline ofanother mammalian species, such as a mouse, have been grafted onto humanframework sequences.

The term “human monoclonal antibody” refers to antibodies displaying asingle binding specificity which have variable regions in which both theframework and CDR regions are derived from human sequences.

The term “recombinant human antibody”, as used herein, includes allhuman antibodies that are prepared, expressed, created or isolated byrecombinant means, such as antibodies isolated from an animal (e.g., amouse) that is transgenic or transchromosomal for human immunoglobulingenes or a hybridoma prepared therefrom, antibodies isolated from a hostcell transformed to express the human antibody, e.g., from atransfectoma, antibodies isolated from a recombinant, combinatorialhuman antibody library, and antibodies prepared, expressed, created orisolated by any other means that involve splicing of all or a portion ofa human immunoglobulin gene, sequences to other DNA sequences. Suchrecombinant human antibodies have variable regions in which theframework and CDR regions are derived from human germline immunoglobulinsequences. In certain embodiments, however, such recombinant humanantibodies can be subjected to in vitro mutagenesis (or, when an animaltransgenic for human Ig sequences is used, in vivo somatic mutagenesis)and thus the amino acid sequences of the V_(H) and V_(L) regions of therecombinant antibodies are sequences that, while derived from andrelated to human germline V_(H) and V_(L) sequences, may not naturallyexist within the human antibody germline repertoire in vivo.

As used herein, “isotype” refers to the antibody class (e.g., IgM, IgA,IgD, IgE and IgG such as IgG1, IgG2, IgG3 or IgG4) that is provided bythe heavy chain constant region genes.

The phrases “an antibody recognizing an antigen” and “an antibodyspecific for an antigen” are used interchangeably herein with the term“an antibody which binds specifically to an antigen”.

As used herein, an antibody that “specifically binds to BAFFRpolypeptide” or an “anti-BAFFR antibody” refers to an antibody thatbinds to human BAFFR polypeptide of SEQ ID NO: 13 with a K_(D) of 100 nMor less, 10 nM or less, 1 nM or less. An antibody that “cross-reactswith an antigen other than BAFFR” refers to an antibody that binds thatantigen with a K_(D) of 0.5×10⁻⁸ M or less, 5×10⁻⁹ M or less, or 2×10⁻⁹M or less. An antibody that “does not cross-react with a particularantigen” is intended to refer to an antibody that binds to that antigen,with a K_(D) of 1.5×10⁻⁸ M or greater, or a K_(D) of 5-10×10⁻⁸ M or1×10⁻⁷ M or greater. In certain embodiments, such antibodies that do notcross-react with the antigen exhibit essentially undetectable bindingagainst these proteins in standard binding assays.

In one embodiment, a high concentration of an anti-BAFFR antibody in theaqueous pharmaceutical composition of the invention is at least 50mg/mL. In one embodiment, a high concentration is at least 100 mg/mL. Inone embodiment, a high concentration is at least 150 mg/mL. In oneembodiment, a high concentration is at least 200 mg/mL. In oneembodiment, a high concentration is at least 250 mg/mL. In oneembodiment, a high concentration is at least 270 mg/mL. In oneembodiment, a high concentration is at least 3 00 mg/mL.

In one embodiment, the aqueous pharmaceutical composition of theinvention comprises between 50 mg/mL and 300 mg/mL of an anti-BAFFRantibody, for example, MOR6654, especially MOR6654B.

In one embodiment, the aqueous pharmaceutical composition of theinvention comprises between 75 mg/mL and 270 mg/mL of an anti-BAFFRantibody, for example, MOR6654, especially MOR6654B.

In one embodiment, the aqueous pharmaceutical composition of theinvention comprises between 100 mg/mL and 250 mg/mL of an anti-BAFFRantibody, for example, MOR6654, especially MOR6654B.

In one embodiment, the aqueous pharmaceutical composition of theinvention comprises between 100 mg/mL and 200 mg/mL of an anti-BAFFRantibody, for example, MOR6654, especially MOR6654B.

In one embodiment, the aqueous pharmaceutical composition of theinvention comprises 150 mg/mL of an anti-BAFFR antibody, for example,MOR6654, especially MOR6654B.

In one embodiment, the aqueous pharmaceutical composition of theinvention comprises 20 mg/mL of an anti-BAFFR antibody, for example,MOR6654, especially MOR6654B.

In one embodiment, the aqueous pharmaceutical composition of theinvention comprises about 50 mg/mL, about 60 mg/mL, about 70 mg/mL,about 80 mg/mL, about 90 mg/mL, about 100 mg/mL, about 110 mg/mL, about120 mg/mL, about 130 mg/mL, about 140 mg/mL, about 150 mg/mL, about 160mg/mL, about 170 mg/mL, about 180 mg/mL, about 190 mg/mL, about 200mg/mL, about 210 mg/mL, about 220 mg/mL, about 230 mg/mL, about 240mg/mL, about 250 mg/mL, about 270 mg/mL or about 300 mg/mL of ananti-BAFFR antibody, for example, MOR6654, especially MOR6654B.

Furthermore, the aqueous pharmaceutical compositions according to theinvention as described herein in the various embodiments are stable suchthat, even after storage for 4 weeks at 2-8° C., less than 5%, 4%, 3%,2%, 1%, 0.05% or 0.01% of the total anti-BAFFR antibody is aggregated asmeasured by SEC-HPLC.

The aqueous pharmaceutical compositions according to the invention asdescribed herein in the various embodiments are stable such that, evenafter storage for 2 month at 2-8° C., less than 5%, 4%, 3%, 2%, 1%,0.05% or 0.01% of the total anti-BAFFR antibody is aggregated asmeasured by SEC-HPLC.

The aqueous pharmaceutical compositions may include, in addition to theanti-BAFFR antibody, further components such as one or more of thefollowing: (i) a stabilizer; (ii) a buffering agent; (iii) a surfactant;and (iv) a free amino acid. Inclusion of each of such additionalcomponents can give compositions with low aggregation of the anti-BAFFRantibody.

Suitable stabilizer for use with the invention can act, e.g., asviscosity enhancing agents, bulking agents, solubilizing agents, and/orthe like. The stabilizer can be ionic or non ionic (e.g. sugars). Assugars they include, but are not limited to, monosaccharides, e.g.,fructose, maltose, galactose, glucose, D-mannose, sorbose and the like;disaccharides, e.g. lactose, sucrose, trehalose, cellobiose, and thelike; polysaccharides, e.g. raffinose, melezitose, maltodextrins,dextrans, starches, and the like; and alditols, such as mannitol,xylitol, maltitol, lactitol, xylitol sorbitol (glucitol) and the like.For example, the sugar may be sucrose, trehalose, raffinose, maltose,sorbitol or mannitol. The sugar may be a sugar alcohol or an aminosugar. Sucrose is particularly useful. As ionic stabilizer they includesalts such as NaCl or amino acid components such as arginine-HCl.

Suitable buffering agents for use with the invention include, but arenot limited to, organic acid salts such as salts of citric acid,ascorbic acid, gluconic acid, carbonic acid, tartaric acid, succinicacid, acetic acid or phtalic acid; Tris, thomethamine hydrochloride, orphosphate buffer. In addition, amino acid components can also be used asbuffering agent. Such amino acid component includes without limitationglycine and histidine. A histidine buffer is particularly useful.

The aqueous pharmaceutical compositions include such buffering agent orpH adjusting agent to provide improved pH control. In one embodiment, anaqueous pharmaceutical composition of the invention has a pH between 5.0and 8.0, between 5.0 and 7.0, between 5.5 and 7.0, or between 5.5 and6.5. In a specific embodiment, an aqueous pharmaceutical composition ofthe invention has a pH of about 6.0.

As used herein, the term “surfactant” refers to organic substanceshaving amphipathic structures; i.e., they are composed of groups ofopposing solubility tendencies, typically an oil-soluble hydrocarbonchain and a water-soluble ionic group. Surfactants can be classified,depending on the charge of the surface-active moiety, into anionic,cationic and dispersing agents for various pharmaceutical compositionsand preparations of biological materials.

Suitable surfactants for use with the invention include, but are notlimited to, non-ionic surfactants, ionic surfactants and zwitterionicsurfactants. Typical surfactants for use with the invention include, butare not limited to, sorbitan fatty acid esters (e.g. sorbitanmonocaprylate, sorbitan monolaurate, sorbitan monopalmitate), sorbitantrioleate, glycerine fatty acid esters (e.g. glycerine monocaprylate,glycerine monomyristate, glycerine monostearate), polyglycerine fattyacid esters (e.g. decaglyceryl monostearate, decaglyceryl distearate,decaglyceryl monolinoleate), polyoxyethylene sorbitan fatty acid esters(e.g. polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitanmonooleate, polyoxyethylene sorbitan monostearate, polyoxyethylenesorbitan monopalmitate, polyoxyethylene sorbitan trioleate,polyoxyethylene sorbitan tristearate), polyoxyethylene sorbitol fattyacid esters (e.g. polyoxyethylene sorbitol tetrastearate,polyoxyethylene sorbitol tetraoleate), polyoxyethylene glycerine fattyacid esters (e.g. polyoxyethylene glyceryl monostearate), polyethyleneglycol fatty acid esters (e.g. polyethylene glycol distearate),polyoxyethylene alkyl ethers (e.g. polyoxyethylene lauryl ether),polyoxyethylene polyoxypropylene alkyl ethers (e.g. polyoxyethylenepolyoxypropylene glycol, polyoxyethylene polyoxypropylene propyl ether,polyoxyethylene polyoxypropylene cetyl ether), polyoxyethylenealkylphenyl ethers (e.g. polyoxyethylene nonylphenyl ether),polyoxyethylene hydrogenated castor oils (e.g. polyoxyethylene castoroil, polyoxyethylene hydrogenated castor oil), polyoxyethylene beeswaxderivatives (e.g. polyoxyethylene sorbitol beeswax), polyoxyethylenelanolin derivatives (e.g. polyoxyethylene lanolin), and polyoxyethylenefatty acid amides (e.g. polyoxyethylene stearic acid amide); C₁₀-C₁₈alkyl sulfates (e.g. sodium cetyl sulfate, sodium lauryl sulfate, sodiumoleyl sulfate), polyoxyethylene C₁₀-C₁₈ alkyl ether sulfate with anaverage of 2 to 4 moles of ethylene oxide units added (e.g. sodiumpolyoxyethylene lauryl sulfate), and C₁-C₁₈ alkyl sulfosuccinate estersalts (e.g. sodium lauryl sulfosuccinate ester); and natural surfactantssuch as lecithin, glycerophospholipid, sphingophospholipids (e.g.sphingomyelin), and sucrose esters of C₁₂-C₁₈ fatty acids. A compositionmay include one or more of these surfactants. Preferred surfactants arepolyoxyethylene sorbitan fatty acid esters e.g. polysorbate 20, 40, 60or 80. Polysorbate 20 (Tween 20) is particularly useful.

Suitable free amino acids for use with the invention include, but arenot limited to, arginine, lysine, histidine, ornithine, isoleucine,leucine, alanine, glycine glutamic acid or aspartic acid. The inclusionof a basic amino acid is preferred i.e. arginine, lysine and/orhistidine. If a composition includes histidine then this may act both asa buffering agent and a free amino acid, but when a histidine buffer isused it is typical to include a non-histidine free amino acid e.g. toinclude histidine buffer and lysine. An amino acid may be present in itsD- and/or L-form, but the L-form is typical. The amino acid may bepresent as any suitable salt e.g. a hydrochloride salt, such asarginine-HCl.

Other contemplated excipients, which may be utilized in the aqueouspharmaceutical compositions of the invention include, for example,flavoring agents, antimicrobial agents, sweeteners, antioxidants,antistatic agents, lipids such as phospholipids or fatty acids, steroidssuch as cholesterol, protein excipients such as serum albumin (humanserum albumin), recombinant human albumin, gelatin, casein, salt-formingcounterions such sodium and the like. These and additional knownpharmaceutical excipients and/or additives suitable for use in theformulations of the invention are known in the art, e.g., as listed in“The Handbook of Pharmaceutical Excipients, 4th edition, Rowe et al.,Eds., American Pharmaceuticals Association (2003); and Remington: theScience and Practice of Pharmacy, 21th edition, Gennaro, Ed., LippincottWilliams & Wilkins (2005).

The aqueous pharmaceutical compositions of the invention may includefurther active ingredients in addition to the anti-BAFFR antibody.Further pharmacological agents may include, for instance,chemotherapeutic compounds.

Target Diseases and Disorders

The aqueous pharmaceutical compositions of the invention comprisinganti-BAFFR antibodies can be used to treat, ameliorate or prevent avariety of diseases or disorders. Pharmaceutical compositions comprisinganti-BAFFR antibodies are particularly useful to treat BAFFR relateddisorders such as autoimmune disorders, e.g., systemic lupuserythematosus, Sjögren's syndrome, Pemphigus vulgaris, rheumatoidarthritis, multiple sclerosis and B cell neoplasms such as acutelymphoblastic leukemia (ALL) and B-cell chronic lymphocytic leukemia(CLL).

As used herein, “a BAFFR-related disorder” includes conditionsassociated with or characterized by aberrant BLyS levels and/or diseasesor conditions that can be treated by depleting or killing B cells. Theseincludes, without limitations, inflammatory conditions, autoimmunediseases, severe infections, and organ or tissue transplant rejection.These further include B-cell neoplasms.

For example, the aqueous pharmaceutical compositions of the inventioncomprising anti-BAFFR antibodies may be used for the treatment,amelioration or prevention of recipients of heart, lung, combinedheart-lung, liver, kidney, pancreatic, skin or corneal transplants,including allograft rejection or xenograft rejection, and for theprevention of graft-versus-host disease, such as following bone marrowtransplant, and organ transplant associated arteriosclerosis. Further,the aqueous pharmaceutical compositions of the invention are useful insolid organ transplantation and in antibody-mediated acute and chronictransplant rejection.

The aqueous pharmaceutical compositions of the invention comprisinganti-BAFFR antibodies are useful for the treatment, prevention, oramelioration of autoimmune disease and of inflammatory conditions, inparticular inflammatory conditions with an etiology including anautoimmune component such as arthritis (for example rheumatoidarthritis, arthritis chronica progrediente and arthritis deformans) andrheumatic diseases, including inflammatory conditions and rheumaticdiseases involving bone loss, inflammatory pain, spondyloarhropathiesincluding ankylosing spondylitis, Reiter syndrome, reactive arthritis,psoriatic arthritis, and enterophathics arthritis, hypersensitivity(including both airways hypersensitivity and dermal hypersensitivity)and allergies. Specific auto-immune diseases for which antibodies of theinvention may be employed include autoimmune haematological disorders(including e.g. hemolytic anaemia, aplastic anaemia, pure red cellanaemia and idiopathic thrombocytopenia), acquired hemophilia A, coldagglutinin disease, cryoglobulinemia, thrombotic thrombocytopenicpurpura, Sjögren's syndrome, systemic lupus erythematosus, inflammatorymuscle disorders, polychondritis, scleroderma, vasculitis such ascryoglobulinemia, large vessel vasculitides such as giant cellarteritis, polymyalgia rheumatica, necrotizing vasculitides, includinganti-neutrophil cytoplasmic antibody-associated vasculitis, Takayasu'sarteritis, polyarteritis nodosa, Henoch-Schonlein purpura, andChurg-Strauss syndrome, IgM mediated neuropathy, seronegativespondarthritis, opsoclonus myoclonus syndrome, Wegener granulomatosis,dermatomyositis, anti-neutrophil cytoplasmatic autoantibody (ANCA)vasculitis, chronic active hepatitis, myasthenia gravis, psoriasis,Steven-Johnson syndrome, pemphigus vulgaris, pemphigus foliacius,idiopathic sprue, autoimmune inflammatory bowel disease (including e.g.ulcerative colitis, Crohn's disease and Irritable Bowel Syndrome),endocrine ophthalmopathy, Graves' disease, sarcoidosis, multiplesclerosis, neuromyelitis optica, primary biliary cirrhosis, juvenilediabetes (diabetes mellitus type I), uveitis (anterior, intermediate andposterior as well as panuveitis), keratoconjunctivitis sicca and vernalkeratoconjunctivitis, interstitial lung fibrosis, psoriatic arthritisand glomerulonephritis (with and without nephrotic syndrome, e.g.including idiopathic nephrotic syndrome or minimal change nephropathy),acute nephritic lupus, tumors, inflammatory disease of skin and cornea,myositis, loosening of bone implants, metabolic disorders, such asatherosclerosis, diabetes, and dislipidemia.

The aqueous pharmaceutical compositions of the invention may also beuseful in preventing, ameliorating or treating B-cell neoplasms.Examples of such diseases and conditions include, but are not limitedto, B-cell Non-Hodgkin's lymphomas, such as small lymphocytic lymphoma,lymphoplasmacytoid lymphoma, mantle cell lymphoma, follicular lymphoma,mucosa-associated lymphoid tissue lymphoma, diffuse large cell lymphoma,and Burkitt's lymphoma; acute lymphoblastic leukemia (ALL), precursorB-lymphoblastic leukemia; B-cell chronic lymphocytic leukemia (CLL), andmultiple myeloma. Other B-cell neoplasms are encompassed within thescope of the invention.

Patient Administration

A pharmaceutical composition of the invention can be administered to apatient. Administration will typically be by infusion or via a syringe.Thus, the invention provides a delivery device (e.g. a syringe)including a pharmaceutical composition of the invention (e.g.,pre-filled syringe). Patients will receive an effective amount of theanti-BAFFR antibody as the principal active ingredient i.e. an amountthat is sufficient to treat, ameliorate, or prevent the disease ordisorder in question. Therapeutic effects may also include reduction inphysical symptoms. The optimum effective amount and concentration ofantibody for any particular subject will depend upon various factors,including the patient's age size health and/or gender, the nature andextent of the condition, the activity of the particular antibody, therate of its clearance by the body, and also on any possible furthertherapeutic(s) administered in combination with the antibody. Theeffective amount delivered for a given situation can be determinedwithin the judgment of a clinician. For purposes of the presentinvention, an effective dose may be from about 0.005 mg/kg to about 50mg/kg, or about 0.05 mg/kg to about 10 mg/kg. Known antibody-basedpharmaceuticals provide guidance in this respect e.g. HERCEPTIN™ isadministered with an initial loading dose of 4 mg/kg body weight and aweekly maintenance dose of 2 mg/kg body weight; RITUXAN™ is administeredweekly at 375 mg/m²; SYNAGIS™ is administered intramuscularly at 15mg/kg body weight; etc.

The invention provides a method for delivering a monoclonal antibody toa mammal, comprising a step of administering to the patient apharmaceutical composition of the invention.

The invention also provides formulations of the invention as describedherein in the various embodiments for use as medicaments e.g. for use indelivering an antibody to a mammal, or for use in treating, preventingor ameliorating one or more of the diseases and disorders describedabove.

The mammal is preferably a human but may also be, for example, a horseor a cow or a dog or a cat. The antibodies will ideally be chosen tomatch the target species e.g. a human antibody for human administration,an equine antibody for horses, a canine antibody for dogs, etc. Ifnative host antibodies are not available then transfer of antibodyspecificity from one species to another can be achieved by transfer ofCDR residues (and typically, in addition, one or more frameworkresidues) from a donor antibody into a recipient framework from the hostspecies e.g. as in humanization. Equinized, bovinized, caninized andfelinized antibodies are known in the art. The antibody will bind toBAFFR from the target species, but it may also cross-react with BAFFRfrom other species.

Dosage can be by a single dose schedule or a multiple dose schedule.

Ingredients for forming compositions of the invention may be supplied inhermetically-sealed containers.

The Anti-BAFFR Antibody

The invention concerns the formulation of anti-BAFFR antibodies and morespecifically MOR6654 and MOR6654B.

One suitable antibody that can be comprised in the pharmaceuticalcompositions of the invention is the human recombinant antibody MOR6654,structurally characterized as further described below. The V_(H) aminoacid sequence of such isolated anti-BAFFR antibody is shown in SEQ IDNO: 1. The V_(L) amino acid sequence of such isolated anti-BAFFRantibody is shown in SEQ ID NO: 2. An example of the full length heavychain amino acid sequence of such isolated anti-BAFFR antibody is shownin SEQ ID NO: 9. An example of the full-length light chain amino acidsequence of such isolated anti-BAFFR antibody is shown in SEQ ID NO: 10.Another example of heavy and light chain amino acid sequences of suchisolated anti-BAFFR antibodies are those encoded by the nucleotidesequences of SEQ ID NO: 11 and SEQ ID NO: 12 respectively. Anotherexample of heavy and light chain amino acid sequences of antibodies arethose encoded by corresponding DNA sequences contained in plasmid pBW510as deposited by Novartis Pharma AG, Forum 1, CH-4002 Basel, Switzerland,at DSMZ, Inhoffenstrasse 7B, 38124 Braunschweig, Germany on Apr. 29,2009 with accession number DSM22542.

Other anti-BAFFR antibodies that can be used for preparing thepharmaceutical compositions of the invention include anti-BAFFRantibodies, with amino acids that have been mutated by amino aciddeletion, insertion or substitution, yet have no more than 1, 2, 3, 4 or5 amino acid deletion, insertion or substitution in either the heavy orlight chain regions described above. In a specific embodiment, suchamino acid changes appear only within the framework and/or constantregions and the CDR regions are 100% identical to the heavy chain CDR1,CDR2 and CDR3 regions of SEQ ID NO: 3, 4 and 5 and to the light chainCDR1, CDR2 and CDR3 regions of SEQ ID NO: 6, 7, and 8 respectively. Inone more specific embodiment, the changes that have been made are onlyconservative amino acid substitutions outside of the CDR regions.

Conservative amino acid substitutions are ones in which the amino acidresidue is replaced with an amino acid residue having a similar sidechain. Families of amino acid residues having similar side chains havebeen defined in the art. These families include amino acids with basicside chains (e.g., lysine, arginine, histidine), acidic side chains(e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g.,glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine,tryptophan), nonpolar side chains (e.g., alanine, valine, leucine,isoleucine, proline, phenylalanine, methionine), beta-branched sidechains (e.g., threonine, valine, isoleucine) and aromatic side chains(e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, one ormore amino acid residues outside of the CDR regions of an anti-BAFFRantibody, can be replaced with other amino acid residues from the sameside chain family, and the altered antibody can be tested for retainedfunction, in particular the same binding properties to BAFFR.

Antibodies may typically be glycosylated. N-linked glycans attached tothe C_(H)2 domain of a heavy chain, for instance, can influence Clq andFcR binding, and aglycosylated antibodies may have lower or differentaffinity for these receptors. The glycan structure can also affectactivity e.g. differences in complement-mediated cell death may be seendepending on the number of galactose sugars (0, 1 or 2) at the terminusof a glycan's biantennary chain. An antibody's glycans preferably do notlead to a human immunogenic response after administration.

Additionally or alternatively, an antibody can be made that has analtered type of glycosylation, such as a hypofucosylated ornon-fucosylated antibody having reduced amounts of or no fucosylresidues or an antibody having increased bisecting GlcNac structures.Such altered glycosylation patterns have been demonstrated to increasethe antibody-dependent cell-mediated cytotoxicity (ADCC) ability ofantibodies. Such carbohydrate modifications can be accomplished by, forexample, expressing the antibody in a host cell with alteredglycosylation machinery. Cells with an altered glycosylation machineryhave been described in the art and can be used as host cells in which toexpress recombinant antibodies of the invention to thereby produce anantibody with altered glycosylation. For example, EP 1,176,195 by Hanget al. describes a cell line with a functionally disrupted FUT8 gene,which encodes a fucosyl transferase, such that antibodies expressed insuch a cell line exhibit hypofucosylation or are devoid of fucosylresidues. Therefore, in one embodiment, the anti-BAFFR antibodies thatare included in the pharmaceutical compositions of the invention areproduced by recombinant expression in a cell line which exhibithypofucosylation or non-fucosylation pattern, for example, a mammaliancell line with deficient expression of the FUT8 gene encodingfucosyltransferase.

As used herein, the term MOR6654 encompasses any type of glycosyationpattern. In a specific embodiment, the pharmaceutical compositionscomprises an anti-BAFFR antibody consisting of MOR6654 as produced in acell line which exhibits a hypofucosylation or non-fucosylation pattern,such as MOR6654B, which exhibit non-fucosylation pattern (devoid offucosyl residues). PCT Publication WO 03/035835 by Presta describes avariant CHO cell line, Lec13 cells, with reduced ability to attachfucose to Asn(297)-linked carbohydrates, also resulting inhypofucosylation of antibodies expressed in that host cell (see alsoShields, R. L. et al., 2002 J. Biol. Chem. 277:26733-26740). PCTPublication WO 99/54342 by Umana et al. describes cell lines engineeredto express glycoprotein-modifying glycosyl transferases (e.g.,beta(1,4)-N acetylglucosaminyltransferase III (GnTIII)) such thatantibodies expressed in the engineered cell lines exhibit increasedbisecting GlcNac structures which results in increased ADCC activity ofthe antibodies (see also Umana et al., 1999 Nat. Biotech. 17:176-180).Eureka Therapeutics further describes genetically engineered CHOmammalian cells capable of producing antibodies with altered mammalianglycosylation pattern devoid of fucosyl residues(http://www.eurekainc.com/about_us/companyoverview.html). Alternatively,the anti-BAFFR antibodies can be produced in yeasts or filamentous fungiengineered for mammalian-like glycosylation pattern and capable ofproducing antibodies lacking fucose as glycosylation pattern (see forexample EP1297172B1).

Another modification of the anti-BAFFR antibodies herein that iscontemplated by the invention is pegylation. An antibody can bepegylated to, for example, increase the biological (e.g., serum)half-life of the antibody. To pegylate an antibody, the antibody, orfragment thereof, typically may be reacted with polyethylene glycol(PEG), such as a reactive ester or aldehyde derivative of PEG, underconditions in which one or more PEG groups become attached to theantibody or antibody fragment. The pegylation can be carried out by anacylation reaction or an alkylation reaction with a reactive PEGmolecule (or an analogous reactive water-soluble polymer).

As used herein, the term “polyethylene glycol” is intended to encompassany of the forms of PEG that have been used to derivatize otherproteins, such as mono (C1-C10) alkoxy- or aryloxy-polyethylene glycolor polyethylene glycol-maleimide. In certain embodiments, the antibodyto be pegylated is an aglycosylated antibody. Methods for pegylatingproteins are known in the art and can be applied to the antibodies ofthe invention. See for example, EP 0 154 316 by Nishimura et al. and EP0 401 384 by Ishikawa et al.

Any other natural or non-natural post-translational modification ofanti-BAFFR antibodies (e.g. MOR6654) is further contemplated as specificembodiments of anti-BAFFR antibodies that could be used for preparingthe pharmaceutical compositions of the invention.

Antibodies can be prepared in a form free from products with which theywould naturally be associated. Contaminant components of an antibody'snatural environment include materials such as enzymes, hormones, orother host cell proteins.

EXAMPLES Preparing Anti-BAFFR Antibodies

Antibody MOR6654 binds specifically to BAFFR and is also described ininternational application published as WO2010/007082. It is a human IgG1kappa antibody obtained via phage display. Its heavy and light chainsconsist of SEQ ID NOs: 9 and 10. The Tables 1 and 2 below summarize thesequence characteristics of MOR6654.

TABLE 1 Brief description of the sequences listed in the sequencelisting of Table 2 SEQ ID NO: Description of the sequence 1 Amino acidsequence of the variable region (V_(H)) of the heavy chain of MOR6654 2Amino acid sequence of the variable region (V_(L)) of the light chain ofMOR6654 3 Amino acid sequence of HCDR1 of MOR6654 4 Amino acid sequenceof HCDR2 of MOR6654 5 Amino acid sequence of HCDR3 of MOR6654 6 Aminoacid sequence of LCDR1 of MOR6654 7 Amino acid sequence of LCDR2 ofMOR6654 8 Amino acid sequence of LCDR3 of MOR6654 9 Amino acid sequenceof the full length heavy chain of MOR6654 10 Amino acid sequence of thefull length light chain of MOR6654 11 Nucleotide sequence encoding SEQID NO: 1 12 Nucleotide sequence encoding SEQ ID NO: 2 13 Human BAFFRamino acid sequence 14 Full length nucleotide sequence (including leadersequence and constant part) of MOR6654 heavy chain; nt 1-57 = leader; nt58-429 = VH; nt 430-1419 = constant region (hIgG1) 15 Full lengthnucleotide sequence (including leader sequence and constant part) ofMOR6654 light chain; nt 1-60 = leader; nt 61-384 = VL; nt 385-705 =constant region (hkappa)

TABLE 2 Sequence listing SEQ ID NO: Amino acid or Nucleotide Sequence  1QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWGWIRQSPGRGLEWLGRIYYRSKWYNSYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCARYDWVPKIGVFDSWGQGTLV TVSS  2DIVLTQSPATLSLSPGERATLSCRASQFISSSYLSWYQQKPGQAPRLLIYGSSSRATGVPARFSGSGSGTDFTLTISSLE PEDFAVYYCQQLYSSPMTFGQGTKVEIK  3GDSVSSNSAAWG  4 RIYYRSKWYNSYAVSVKS  5 YDWVPKIGVFDS  6 RASQFISSSYLS  7GSSSRAT  8 QQLYSSPMT  9 QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWGWIRQSPGRGLEWLGRIYYRSKWYNSYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCARYDWVPKIGVFDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPGK 10DIVLTQSPATLSLSPGERATLSCRASQFISSSYLSWYQQKPGQAPRLLIYGSSSRATGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQLYSSPMTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC 11CAGGTGCAGCTGCAGCAGAGCGGCCCAGGCCTGGTCAAGCCCTCTCAGACCCTGTCACTGACCTGCGCCATTTCAGGCGACAGCGTGAGCAGCAACAGCGCCGCCTGGGGCTGGATCAGGCAGAGCCCCGGTAGGGGCCTGGAATGGCTGGGCAGGATCTACTACAGGTCCAAGTGGTACAACAGCTACGCCGTGAGCGTGAAGAGCAGGATCACCATCAACCCTGACACCAGCAAGAACCAGTTCTCACTGCAGCTCAACAGCGTGACCCCCGAGGACACCGCCGTGTACTACTGCGCCAGATACGACTGGGTGCCCAAGATCGGCGTGTTCGACAGCTGGGGCCAGGGCACCCTGGTG ACCGTGTCAAGC 12GATATCGTGCTGACACAGAGCCCCGCCACCCTGAGCCTGAGCCCAGGCGAGAGGGCCACCCTGTCCTGCAGGGCCAGCCAGTTTATCAGCAGCAGCTACCTGTCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTAGACTGCTGATCTACGGCAGCTCCTCTCGGGCCACCGGCGTGCCCGCCAGGTTCAGCGGCAGCGGCTCCGGCACCGACTTCACCCTGACAATCAGCAGCCTGGAGCCCGAGGACTTCGCCGTGTACTACTGCCAGCAGCTGTACAGCTCACCCATGACCTTCGGCCAGGGCACCAAGGTGGAGAT CAAG 13MRRGPRSLRGRDAPAPTPCVPAECFDLLVRHCVACGLLRTPRPKPAGASSPAPRTALQPQESVGAGAGEAALPLPGLLFGAPALLGLALVLALVLVGLVSWRRRQRRLRGASSAEAPDGDKDAPEPLDKVIILSPGISDATAPAWPPPGEDPGTTPPGHS VPVPATELGSTELVTTKTAGPEQQ 14ATGGCCTGGGTGTGGACCCTGCCCTTCCTGATGGCCGCTGCCCAGTCAGTGCAGGCCCAGGTGCAGCTGCAGCAGAGCGGCCCAGGCCTGGTCAAGCCCTCTCAGACCCTGTCACTGACCTGCGCCATTTCAGGCGACAGCGTGAGCAGCAACAGCGCCGCCTGGGGCTGGATCAGGCAGAGCCCCGGTAGGGGCCTGGAATGGCTGGGCAGGATCTACTACAGGTCCAAGTGGTACAACAGCTACGCCGTGAGCGTGAAGAGCAGGATCACCATCAACCCTGACACCAGCAAGAACCAGTTCTCACTGCAGCTCAACAGCGTGACCCCCGAGGACACCGCCGTGTACTACTGCGCCAGATACGACTGGGTGCCCAAGATCGGCGTGTTCGACAGCTGGGGCCAGGGCACCCTGGTGACCGTGTCAAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCAGAGCTGCTGGGCGGACCCTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAATACAAGTGCAAGGTCTCCAACAAGGCCCTGCCAGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCTCCCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCT GAGCCTGTCCCCCGGCAAG 15ATGAGCGTGCTGACCCAGGTGCTGGCTCTGCTGCTGCTGTGGCTGACCGGCACCAGATGCGATATCGTGCTGACACAGAGCCCCGCCACCCTGAGCCTGAGCCCAGGCGAGAGGGCCACCCTGTCCTGCAGGGCCAGCCAGTTTATCAGCAGCAGCTACCTGTCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTAGACTGCTGATCTACGGCAGCTCCTCTCGGGCCACCGGCGTGCCCGCCAGGTTCAGCGGCAGCGGCTCCGGCACCGACTTCACCCTGACAATCAGCAGCCTGGAGCCCGAGGACTTCGCCGTGTACTACTGCCAGCAGCTGTACAGCTCACCCATGACCTTCGGCCAGGGCACCAAGGTGGAGATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGAC CAAGAGCTTCAACAGGGGCGAGTGC

Examples of Formulations

A high concentration liquid formulation of MOR6654B was desired and soformulation studies were performed. A liquid formulation comprising asugar, a buffering agent and a surfactant was stable and could maintainhigh antibody concentrations.

The antibody may be produced in mammalian host cells, such as, a CHOcell line transfected with expression vectors carrying heavy and lightchain coding sequences under suitable expression promoters.

The antibody is preferably produced in a mammalian cell line, e.g. a CHOcell line, modified by using, for example, the Potelligent™ technology(BioWa, Inc.) leading to deficient expression of the FUT8 gene encodingfucosyltransferase. The resulting antibody is non-fucosylated anddesignated herein as MOR6654B.

The development of a liquid MOR6654B formulation in a vial or apre-filled syringe consisted of two studies. A first screen was made todefine the excipients in the formulation. In a second screen theselected formulation was confirmed in the final primary packaging.

Stability and Analytical Plan

The following analytical methods were performed: UV Assay, SizeExclusion Chromatography HPLC, Dynamic Light Scattering, CationicExchange Chromatography HPLC, Reverse Phase Chromatography HPLC, ALPAnalyzer, Turbidity, pH value, osmolality, MFI (Micro-Flow Imaging),viscosity, color, visual inspection.

Stability Determination

Stability was further determined after subjecting the formulations tocertain stress conditions including agitation, freeze-thaw cycles andprolonged exposure to light (1 month at 40° C.).

Turbidimetric Method

Turbidity was measured by a turbidimetric method.

The turbidimeter measurements for the samples were performed byfollowing the operational manual (Model 2100AN Instrument Manual,Number: 47901-88, November 2006, Edition 2). At the start of theanalysis the calibration curve for the instrument was checked byanalyzing 5 calibration samples, including water. If the values measuredby the instrument were within 5% of the standards value, than thecalibration curve passed. If any of the standards failed the 5%acceptance criteria, than the instrument was recalibrated following theoperational manual. After the instrument passed calibration, the sampleswere loaded in 11 mm flat bottom text tubes and analyzed.

Size Exclusion Chromatographic (SEC) Method

In the SEC method used to analyze the liquid formulations, the followingmethod parameters were used:

Column: TSKgel G3000SWXL Analysis Buffer: 150 mM K-phosphate, pH6.5+/−0.1

Flow Rate: 0.4 mL/minColumn temperature: 30° C.

Detection: 210 nm

Injection volume: 10 ulSample temperature: approx. 5° C.Run time: 40 minutesData sampling rate: 1.0 points/second (Chromeleon step=1.0)

Cation Exchange Chromatography (CEX) HPLC Method

The first step for sample analysis using the CEX HPLC method was todilute all formulations to 10 mg/mL using water. The second step wastaking the 10 mg/mL diluted sample and diluting again with mobile phaseA to 3 mg/mL. The sample was then loaded into the HPLC and analyzed.

Mobile Phase A: 25 mM sodium phosphate, pH 6.5Mobile Phase B: 25 mM sodium phosphate, 250 mM NaCl, pH 6.5Flow Rate: 1.0 mL/min

Column Temperature: 30° C.+/−2° C. Sample Temperature: 5° C.+/−2° C.Injection Volume: 40 ul

Run Time (min): 60

Detection: 215 nm Gradient:

Time % A % B 0 100 0 46 0 100 51 0 10 52 100 0 60 100 0

RP HPLC Method

The RP HPLC method used to analyze the LB-120 samples was performed byfollowing the following RP method parameters. The gradient in thetesting protocol resulted in the protein being eluded in the column voidvolume.

Method Parameters

-   -   Column Information: PoroShell 300SB-C8    -   Mobile Phase A: 90% (v/v) H2O/10% (v/v) ACN/0.1% (v/v) TFA    -   Mobile Phase B: 10% (v/v) H2O/90% (v/v) ACN/0.1% (v/v) TFA    -   Flow rate: 2 mL/min    -   Column temperature: 80° C.    -   Detection: 210 nm    -   Injection volume: 5 μL    -   Sample temperature: Approx. 5° C.    -   Run time: 6 minutes    -   Gradient:

Time % A % B 0 90 10 5 75 25 5.1 90 10 6 90 10

MFI Method

All of the stability samples were tested using a Protein SimpleMicro-Flow Imaging (MFI) instrument, model DPA 4200 with a standard 100μm flow cell (1.6 mm, SP3 with Silane coating, cat#4002-002-001) and a5× objective. The software version for MFI View System Software was2-R2.6.2.21.2171.

The instrument set-up and selected features included:

-   -   Edge Particle Rejection—selected    -   Fill Particles—selected (except TO)    -   Sample Volume: 0.5 mL    -   Purge Volume: 0.15 mL    -   Approximate Sample Volume: 0.30 mL (auto calculated)

On the day of testing, either three (single testing) or four (duplicatetesting) syringes per each condition were pooled into individual 15 mLNunc tubes (Catalogue#339651). All work was performed in a laminar flowhood and MFI samples were tested undiluted.

Run sequences consisted of running water blanks prior to each sample andstandard to ensure background levels of particles were with reason(typically below 600 particles/mL or less than 1% of sampleparticles/mL). Water flushes of 30 mL or more were used betweendifferent samples and between samples and standards prior to measuringbackground particle levels (blanks). If blanks were not acceptableadditional flushing was performed and another blank was run. Waterflushes of 10 mL were used between replicate samples and replicatestandards to adequately clear the system of air bubbles and reduceparticles. Millipore Direct-Q type 1 water was used for blanks andflushing (0.22 μm filtered, 18.2 MΩ quality). Neptune Barrier 1000 μlTips used to deliver sample to the sample inlet port.

Each sample sequence contained a number of NIST (National Institute ofStandard and Technology) certified 5.0 μm particle standards containing3000 particles/mL greater than or equal to 3 μm in size (Cat# CC05, Lot39588, Exp. July 2012) to determine reproducibility during the run. Theperformance of the standards is listed in Table 3.

TABLE 3 Analysis summary of NIST particle standards using MFI Blank Std3 Std # of Timepoint Average Average Deviations Stds Lot# T0 207 3105±16% 4 39588, bottle 2 T1 304 2222  ±6% 4 39588, bottle 2 T2 337 3109±16% 7 39588, 042512

Individual particle size was determined by using Protein Simplesmeasurement technique known as Equivalent Circular Diameter (ECD). TheECD of an object is expressed in microns and represents the diameter ofa sphere that occupies the same two-dimensional surface area as theparticle. The MFI product platform converts the area of an object intoan ECD value using proprietary conversion techniques to avoid the errorinherent with performing direct calculations based on field of viewdimensions. The conversion techniques are based on a mapping of theentire instrument size range using NIST traceable polystyrene beads.Although the conversion is based on polystyrene beads, the vendors claimis that the unique operation of the MFI product platform will guaranteethe results obtained from the instrument are insensitive to the particlematerial properties. Therefore, the instrument does not requirecalibration against specific sample types for proper operation.

Colorimetric Method

Sample color was analyzed using a standard testing procedure. The colorvalue for each sample is recorded in the European Pharmacopoeia (EP)color definition (Table 4). An example, B1 to B9 is the brown colorscale as defined in the European Pharmacopoeia.

TABLE 4 Color Range for the European Pharmacopoeia From To DescriptionY7 Y1 Yellow Color Scale B9 B1 Brown Color Scale BY7 BY1 Brown-YellowColor Scale GY7 GY1 Green-Yellow Color Scale R7 R1 Red Color Scale

Photostability Testing

Syringes containing each of the four formulations F1-F4 as described inTable 16 were subjected to photostability testing. Overall, twelvesyringes (three each per formulation) were placed on the surface of abox covered with white printer paper 3.5 inches below the lamp source.Syringes were placed in numeric order as follows (1 2 3 4 1 2 3 4 1 2 34) about 1 inch apart. The end syringes (1 and 4) were each 5 inchesfrom the ends of the lamp.

The light sources were two cool-white fluorescence lamps (GE EcoluxF20T12-CW-ECO, 20 W each) and two near-UV fluorescent lamps withspectral distributions from 320 to 400 nm, 20 W each. The exposure timeat ambient temperature was 14 days for the cool-white fluorescent lampsand 88 hours for the near-UV lamps.

Agitation Studies

Three syringes for each of the different formulations to be tested wereremoved from refrigeration conditions and pooled in 15 mL Nunc conicaltubes and individually secured to the platform of a Thermo ScientificMaxQ 2000 orbital shaker and agitated at 150 rpm for 24 hours underambient light and temperature conditions.

Freeze-Thaw (F/T) Cycling Studies

Syringes for each of the different formulations to be tested, wereremoved from refrigeration conditions and pooled in 15 mL Nunc conicaltubes. Pooled samples were then subjected to 5 cycles of freezing (−20°C.) and thawing (using ambient temperature water).

STUDY I: First Liquid Formulations Screen

An initial formulation screen for a liquid formulation of MOR6654B wasset up testing e.g. different buffers, stabilizers, excipients and pHvalues. A few formulations were also investigated to gain experience forthe primary packing selection and to gain experience on the stability ofMOR6654B liquid formulation in the different types of PFS (Pre filledsyringes).

Preparation of Samples

The formulations were produced using Drug Substance (MOR6654B) obtainedfrom a CHO cell line expressing the afucosylated monoclonal antibodyup-concentrated to 160 g/L in water. MOR6654B Drug Substance was mixedwith an appropriate amount of excipient dilution solution, sterilefiltered, filled aseptically into sterile 1 mL PFS (0.7 mL fillingvolume) or 6 mL glass vials (3.6 mL fill volume) and stoppered withDaikyo D21-7S V10-F7-3WRS RB2-TR lyo stoppers. All the formulationstested were at 100 g/L of MOR6654B.

TABLE 5 List of formulations, First liquid formulation screen ofMOR6654B (100 g/L) PP (Primary Packaging); HPbCD(hydroxyproyl-b-cyclodextrin) buffer Form pH 20 mM stabilizer surfactantPP 1 6.5 Histidine Trehalose 270 mM Polysorbate 20 0.04% PFS 2 6.5Citrate ArgHCl 150 mM Poloxamer 188 0.3% PFS 3 6.5 Histidine Trehalose270 mM Polysorbate 20 0.04% PFS 4 7 Succinate ArgHCl 150 mM Polysorbate20 0.04% PFS 5 7 Citrate Trehalose 270 mM None PFS 6 7 HistidineMannitol 270 mM Poloxamer 188 0.3% PFS 7 6 Citrate Mannitol 270 mMPolysorbate 20 0.04% PFS 8 6.5 Histidine Trehalose 270 mM Polysorbate 200.04% PFS 9 6 Histidine ArgHCl 150 mM None PFS 10 6 Succinate Trehalose270 mM Poloxamer 188 0.3% PFS 11 6.5 Succinate Mannitol 270 mM None PFS12 6.5 Succinate Trehalose 270 mM Polysorbate 20 0.04% PFS 13 6.5Histidine Trehalose 270 mM HPbCD 2.5 mM PFS 14 6.5 Histidine NaCl 150 mMPolysorbate 20 0.04% PFS 15 6.5 Histidine Trehalose 270 mM Polysorbate20 0.04% VIAL 16 6.5 Histidine Trehalose 270 mM Poloxamer 188 0.3% VIAL

Results Tables of Results for Size Exclusion Chromatography (SEC-HPLC)

TABLE 6 Purity by SEC samples in freeze thaw stress Aggregation MainDegradation Formulation products peak products 1 0.52 99.38 0.10 2 0.5999.31 0.10 3 0.51 99.39 0.10 4 0.62 99.29 0.08 5 0.62 99.29 0.10 6 1.9297.94 0.13 7 1.26 98.66 0.08 8 0.53 99.36 0.10 9 0.56 99.35 0.10 10 0.6499.28 0.08 11 1.75 98.15 0.10 12 0.56 99.38 0.07 13 0.51 99.40 0.09 140.72 99.19 0.08 15 0.56 99.35 0.09 16 0.56 99.36 0.09

TABLE 7 Purity by SEC samples in shaking stress Aggregation MainDegradation Formulation products peak products 1 0.51 99.37 0.12 2 0.6099.31 0.09 3 0.52 99.38 0.10 4 0.68 99.23 0.09 5 0.73 99.16 0.11 6 0.7099.18 0.12 7 0.65 99.25 0.10 8 0.54 99.35 0.11 9 0.53 99.37 0.10 10 0.6499.27 0.09 11 0.64 99.25 0.10 12 0.58 99.30 0.13 13 0.53 99.37 0.10 140.62 99.25 0.13 15 0.57 99.31 0.12 16 0.58 99.30 0.12

TABLE 8 Purity by SEC samples in thermal stress (40° C.) AggregationMain Degradation Formulation products peak products 1 1.09 95.84 3.07 21.05 95.93 3.02 3 1.13 95.72 3.15 4 1.34 95.22 3.44 5 2.06 94.06 3.88 61.58 94.77 3.65 7 1.33 95.75 2.92 8 1.13 95.75 3.12 9 0.90 95.89 3.21 101.24 95.78 2.98 11 1.45 95.48 3.07 12 1.14 95.81 3.05 13 1.02 95.90 3.0814 1.21 95.63 3.17 15 1.32 94.75 3.92 16 1.51 93.56 4.93

TABLE 9 Purity by SEC for t0, and stability samples at 5° C. AggregationAggregation Degradation Degradation Aggregation products productsDegradation products products products 3 months at 6 months at products3 months at 6 months at Form T0 5° C. 5° C. T0 5° C. 5° C. 1 0.46 0.530.54 0.1 0.09 0.12 2 0.54 0.62 0.61 0.09 0.07 0.11 3 0.46 0.56 0.53 0.090.09 0.12 4 0.61 0.77 0.76 0.08 0.07 0.12 5 0.59 0.79 0.76 0.1 0.08 0.136 0.63 0.75 0.69 0.08 0.08 0.14 7 0.59 0.67 0.57 0.08 0.05 0.11 8 0.50.59 0.48 0.1 0.08 0.12 9 0.5 0.57 0.47 0.1 0.08 0.12 10 0.6 0.67 0.510.08 0.06 0.12 11 0.57 0.71 0.55 0.09 0.08 0.12 12 0.52 0.61 0.45 0.090.06 0.13 13 0.51 0.57 0.42 0.11 0.05 0.11 14 0.55 0.65 0.49 0.07 0.040.13 15 0.51 0.61 0.43 0.1 0.09 0.13 16 0.53 0.64 0.44 0.09 0.07 0.14

TABLE 10 Purity by SEC for t0, and stability samples at 25° C.Aggregation Aggregation Degradation Degradation Aggregation productsproducts Degradation products products Form products 3 months at 6months at products 3 months at 6 months at No. T0 25° C. 25° C. T0 25°C. 25° C. 1 0.46 0.8 0.56 0.1 1.41 2.47 2 0.54 0.91 0.54 0.09 1.37 2.3 30.46 0.78 0.58 0.09 1.26 2.44 4 0.61 1.12 0.81 0.08 1.43 2.62 5 0.591.39 1.19 0.1 1.6 2.73 6 0.63 1.1 0.83 0.08 1.42 2.7 7 0.59 0.97 0.570.08 1.23 2.18 8 0.5 0.79 0.52 0.1 1.34 2.36 9 0.5 0.75 0.29 0.1 1.272.23 10 0.6 0.92 0.5 0.08 1.26 2.22 11 0.57 1.06 0.67 0.09 1.31 2.23 120.52 0.84 0.54 0.09 1.28 2.39 13 0.51 0.84 0.47 0.11 1.32 2.33 14 0.550.94 0.51 0.07 1.4 2.38 15 0.51 0.94 0.77 0.1 1.55 3.32 16 0.53 1.080.93 0.09 1.82 3.42

TABLE 11 Charge variants by CEX samples in freeze thaw stress AcidicMain Basic Formulation variants peak variants 1 5.18 65.42 29.40 2 5.1066.21 28.69 3 4.97 66.01 29.01 4 5.27 65.34 29.40 5 5.35 64.79 29.86 65.23 65.78 28.99 7 5.38 65.60 29.02 8 5.24 67.21 27.55 9 5.02 64.2430.74 10 5.56 65.18 29.26 11 5.50 65.04 29.45 12 5.19 66.32 28.49 135.45 65.06 29.49 14 5.43 66.04 28.54 15 5.33 66.18 28.49 16 5.24 66.1128.65

TABLE 12 Charge variants by CEX samples in shaking stress Acidic MainBasic Formulation variants peak variants 1 5.09 66.35 28.56 2 5.03 66.8328.13 3 5.05 66.98 27.97 4 5.07 66.60 28.33 5 5.17 65.47 29.36 6 5.3466.70 27.97 7 5.36 66.63 28.01 8 5.11 66.95 27.94 9 4.78 66.45 28.77 104.99 67.25 27.76 11 5.12 66.21 28.67 12 5.21 67.18 27.60 13 5.41 66.4028.19 14 5.05 67.29 27.66 15 5.14 66.70 28.16 16 5.15 66.84 28.01

TABLE 13 Charge variants by CEX samples in thermal stress (40° C.)Acidic Main Basic Formulation variants peak variants 1 13.41 59.24 27.352 12.00 61.63 26.37 3 14.43 58.54 27.03 4 13.59 60.20 26.21 5 14.4058.18 27.42 6 15.36 58.63 26.01 7 14.83 58.99 26.18 8 14.10 59.34 26.559 12.70 58.92 28.38 10 15.17 58.64 26.19 11 14.09 58.86 27.05 12 14.3159.15 26.54 13 14.54 58.97 26.49 14 12.22 61.15 26.62 15 15.01 58.2126.79 16 16.54 56.43 27.03

TABLE 14 Charge variants by CEX for t0, and stability samples at 5° C.Acidic Acidic Basic Basic Acidic variants variants Basic variantsvariants Form variants 3 months at 6 months at variants 3 months at 6months at No. T0 5° C. 5° C. T0 5° C. 5° C. 1 4.19 6.13 14.43 30.5625.39 20.14 2 4.73 5.66 14.35 30.18 26.33 20.81 3 4.73 6.33 14.9 29.4826.66 20.48 4 4.76 5.71 14.08 29.71 26.16 21.53 5 4.8 5.91 14.47 30.6927.66 20.82 6 4.97 6.22 15.73 29.1 26.14 21.2 7 4.68 5.5 14.33 29.1525.69 21.45 8 4.75 5.99 14.98 29.12 25.91 20.77 9 4.71 5.54 14.13 30.4825.89 21.24 10 4.78 5.83 14.55 29.38 26.11 21.12 11 4.89 5.82 14.0430.24 26.39 21.3 12 4.74 6.2 14.52 28.29 25.8 20.98 13 5.04 6.26 16.1330.22 25.75 21.43 14 5.02 5.7 14.62 28.86 25.26 21.38 15 5.02 5.9 16.8729.02 25.81 22.59 16 5.17 6.01 16.92 29.07 25.48 22.52

TABLE 15 Charge variants by CEX for t0, and stability samples at 25° C.Acidic Acidic Basic Basic Acidic variants variants Basic variantsvariants Form variants 3 months at 6 months at variants 3 months at 6months at No. T0 25° C. 25° C. T0 25° C. 25° C. 1 4.19 7.75 27.71 30.5624.44 21.16 2 4.73 6.81 22.26 30.18 25.64 20.38 3 4.73 6.81 24.95 29.4824.38 20.28 4 4.76 8.02 24.73 29.71 24.96 20 5 4.8 7.67 28.55 30.6923.86 21.46 6 4.97 7.36 25.94 29.1 24.86 18.62 7 4.68 7.05 31.1 29.1523.96 20.23 8 4.75 7.85 27.67 29.12 24.73 20.93 9 4.71 7.72 29.52 30.4824.65 19.3 10 4.78 7.87 27.61 29.38 24.76 20.33 11 4.89 8.7 36.3 30.2423.58 19.85 12 4.74 7.24 25.53 28.29 24.91 20.58 13 5.04 7.41 24.7730.22 24.14 19.91 14 5.02 7.75 29.63 28.86 25.37 21.95 15 5.02 8.09 28.729.02 24.56 20.01 16 5.17 8.18 32.26 29.07 24.94 22.16

Summary of Results, First Liquid Formulation Screen

Sixteen formulations of the monoclonal antibody, MOR6654B, were placedon stability for up to six months at 5° C. and 25° C. In addition, thesame formulations were subjected to agitation stress, repeated F/Tcycling and thermal stress (1 month at 40° C.). A wide variety ofbiophysical and biochemical analytical methods was used to determine ifthere were differences between the formulations in terms of stability.SEC data pointed to a difference in stability among the formulationstested. In the shaking stress samples, formulation 5 (pH 7 withoutsurfactant) showed a higher increase in aggregation products, provingthe beneficial effect of polysorbate. More severe increase inaggregation product was recorded upon freeze thaw stress in the mannitoland sodium chloride formulations.

The outcome of this first study can be summarized as follows: Thepreferred pH is 6.0. The non-ionic stabilizer tested, Trehalose, isbeneficial to the formulation stability. Polysorbate 20 is beneficial tothe stability of the formulation preventing formation of aggregates.

STUDY II: Second Liquid Formulation Screen Formulation of Anti-BAFFRAntibodies

Three formulations (F1, F2, and F3) of MOR6654B with a high antibodyconcentration of 150 mg/mL and one formulation with a lower antibodyconcentration of 20 mg/mL (F4) were evaluated for stability. The fourformulations F1, F2, F3 and F4 were filled into a 1.0 mL siliconizedglass syringe and included buffer, sugar, surfactant and free amino acidas shown in Table 16.

TABLE 16 Composition of experimental formulations Amino MOR6654B BufferSugar Surfactant acid F1 150 mg/mL 20 mM 220 mM 0.04% — histidine; 6.0sucrose polysorbate 20 F2 150 mg/mL 20 mM 220 mM 0.04% — histidine; 6.0trehalose polysorbate 20 F3 150 mg/mL 20 mM 120 mM 0.04% 50 mMhistidine; 6.0 sucrose polysorbate 20 arginine- HCl F4  20 mg/mL 20 mM220 mM 0.04% — histidine; 6.0 sucrose polysorbate 20

Samples of each formulation were prepared and tested for stability atthree different temperatures. The four liquid formulations were placedunder experimental storage conditions and tested for stability followingstorage at

2° C.−8° C. at time points 0 (t₀), and 2 month (t₂)25° C. at time points 0 (t₀) and 2 month (t₂)40° C. at time points 0 (t₀), 1 (t₁) and 2 month (t₂)

In addition to storage at various temperatures, the formulations weresubjected to a number of stress conditions, such as prolonged lightexposure, agitation and multiple freeze-thaw (F/T) cycles. Each samplewas analyzed using a variety of methods.

Results and Discussion, Second Formulation Screen

All four formulations were tested at the same time, depending on thestress condition. At time point zero (t0), measurements were made induplicate, as were the measurements made after two months (t2). Allother samples were only analyzed with single replicates. The morerelevant results are reported below.

Nephelometry

Nephelometry is a turbidometric method used to detect the presence ofsoluble aggregates or to indicate opalescence. The output is listed interms of nephelometric turbidity units (NTUs).

TABLE 17 Nephalometric turbidity units (NTUs)) for MOR6654B formulationsstored at 40° C. for up to two months. t₀ 40° C. for 1 month 40° C. for2 month F1 5.16/5.26 6.19 4.00/4.07 F2 4.63/4.39 4.99 4.12/3.95 F38.94/8.54 9.60 8.34/8.33 F4 3.54/3.63 4.39 3.40/3.39

The nephelometry results show that the protein is quite physicallystable, with no apparent change even after two months at 40° C.

Not surprising, storage at lower temperatures did not produce anyappreciable change in the NTU levels for any of the formulations either(Table 18). Why formulation 3 exhibits a higher number of NTUs is notclear.

TABLE 18 Nephelometric turbidity units (NTUs)) for MOR6654B formulationsstored at 4° C. or 25° C. for two months. t₀ 2-8° C. for 2 month 25° C.for 2 month F1 5.16/5.26 4.56/4.48 4.10/3.65 F2 4.63/4.39 5.56/4.093.43/3.39 F3 8.94/8.54 8.11/7.95 9.57/8.55 F4 3.54/3.63 3.43/3.472.98/2.76

TABLE 19 Nephelometric turbidity units (NTUs)) for MOR6654B formulationssubjected to agitation stress (agit), multiple F/T cycles (F/T), andprolonged exposure to light (photo). t₀ Agit F/T Photo F1 5.16/5.26 6.726.28 4.02 F2 4.63/4.39 5.22 5.26 3.91 F3 8.94/8.54 9.58 9.18 8.59 F43.54/3.63 4.48 4.22 3.15

Finally, nephelometry of the samples subjected to stress conditions,such as agitation, multiple F/T cycles, and prolonged exposure to lightrevealed only small changes in any of the formulations for any of thestress conditions. For example, formulation 1 showed an increase ofabout 1.5 NTU upon agitation and approx. 1 NTU upon F/T stress. Bycomparison, the increase in formulation 2 was less than 1 NTU for eitherstress condition. Likewise, the lower concentration formulation 4, whichalso contains sucrose, shows a slight rise of about 1 NTU uponagitation.

High Pressure Liquid Chromatography (HPLC) Measurements Size ExclusionChromatograph (SEC)

Three kinds of HPLC analyses were performed for these samples, startingwith Size Exclusion Chromatography (SEC). There is a clear loss ofmonomer as measured by SEC for samples stored at 40° C. for one or twomonths (Table 20).

TABLE 20 Purity (in terms of percent monomer) by SEC-HPLC for MOR6654Bsamples stored at 40° C. for up to two months. t₀ 40° C. for 1 month 40°C. for 2 month F1 99.74/99.78 92.71 92.08/92.27 F2 99.64/99.60 93.5192.39/92.61 F3 99.77/99.76 93.26 92.43/92.32 F4 99.67/99.53 93.8093.58/92.87

There is a loss of about 7%-8% monomer as measured by SEC for samplesstored at 40° C. for one or two month.

TABLE 21 Purity (in terms of percent monomer) by SEC- HPLC SEC forMOR6654B samples stored at 4° C. and 25° C. for two months. t₀ 2-8° C.for 2 month 25° C. for 2 month F1 99.74/99.78 99.62/99.63 99.35/99.58 F299.64/99.60 99.61/99.59 99.50/99.35 F3 99.77/99.76 99.58/99.5799.18/99.26 F4 99.67/99.53 99.60/99.57 99.26/99.42

For samples stored at lower temperature, there is no loss of monomer at4° C. and only a small loss of approx. 1% at 25° C. When considering theeffects of the stress conditions, there is a slight decrease in allformulations upon agitation and F/T cycling, but no real differencebetween formulations. Upon photolysis, there is also a small decrease inmonomer content. Again, differences between formulations are not readilyapparent.

TABLE 22 Nephelometric turbidity units (NTUs) for MOR6654B formulationssubjected to agitation stress (agit), multiple F/T cycles (F/T), andprolonged exposure to light (photo). t₀ Agit F/T Photo F1 99.74/99.7899.29 98.86 98.94 F2 99.64/99.60 99.04 98.96 98.86 F3 99.77/99.76 98.8598.98 98.44 F4 99.67/99.53 99.01 98.93 98.68

While an examination of the overall monomer content is helpful, it maybe more useful to consider the overall stability profile. The followingtables have been prepared summarizing the percent of each peak observedat each site. For convenience, the peaks are ordered by their relativeretention time (RRTs) to remove slight variations in run times.

TABLE 23 Overall stability profile as measured by SEC for MOR6654Bsamples stored at 40° C. for up to two months. Time RRT Formulation RepPoint 0.81 0.84 0.88 0.91 0.95 1.00 1.07 1.22 1.38 1.58 Formulation 1 10 0.13 99.74 0.04 0.09 2 0 0.14 99.78 0.03 0.04 Formulation 2 1 0 0.3199.64 0.05 2 0 0.33 99.60 0.07 Formulation 3 1 0 0.17 99.78 0.05 0.05 20 0.19 99.77 0.05 Formulation 4 1 0 0.12 99.71 0.17 2 0 0.17 99.59 0.070.17 Formulation 1 1 1 1.12 92.85 5.35 0.68 Formulation 2 1 1 1.10 93.984.43 0.49 Formulation 3 1 1 0.90 0.07 93.61 4.84 0.58 Formulation 4 1 10.78 94.70 4.03 0.49 Formulation 1 1 2 0.16 0.18 92.08 6.40 1.18 2 20.17 0.21 92.27 6.31 1.05 Formulation 2 1 2 0.44 92.39 6.05 1.12 2 20.19 0.25 92.61 5.94 1.01 Formulation 3 1 2 0.14 0.20 92.43 6.27 0.96 22 0.17 0.19 92.32 6.27 1.05 Formulation 4 1 2 0.30 0.18 93.58 4.79 1.152 2 0.29 0.18 92.87 5.39 1.26

There was no high molecular weight aggregate peak detected at t0 (Table23), and only three impurities found, including on peak at RRT 0.88,which is presumably an oligomer of some type. Upon storage at 40° C. forone month or two months, there is a substantial decrease in monomercontent, with most of the degradation products eluting later than themain peak (Table 23). This indicates that fragmentation is moreproblematic than aggregation for these formulations.

For samples heated at 25° C. for two months, there is very little lossin monomer (<1%) (Table 24). This suggests that the apparent activationenergy for the primary degradation pathway is quite large, which wouldbe consistent with hydrolytic degradation. The same two fragmentationpeaks are seen at RRT 1.07 and 1.22, with the peak at 1.07 being quite abit larger in all samples. In fact, the RRT 1.22 is so small informulations 1 and 2, that they were not integrated in the runs.Otherwise, there are only the smallest differences between formulationsstored at 25° C. when assayed by SEC.

TABLE 24 Overall stability profile as measured by SEC for MOR6654Bsamples stored at 25° C. for two months. Formulation Time RRT 25 C. RepPoint 0.81 0.88 0.91 0.96 1.00 1.07 1.22 1.38 1.58 Formulation 1 1 00.13 99.74 0.04 0.09 2 0 0.14 99.78 0.03 0.04 Formulation 2 1 0 0.3199.64 0.05 2 0 0.33 99.60 0.07 Formulation 3 1 0 0.17 99.78 0.05 0.05 20 0.19 99.77 0.05 Formulation 4 1 0 0.12 99.71 0.17 2 0 0.17 99.59 0.070.17 Formulation 1 1 2 0.33 99.35 0.32 2 2 0.13 99.58 0.29 Formulation 21 2 0.29 99.50 0.21 2 2 0.34 99.35 0.31 Formulation 3 1 2 0.47 99.180.31 0.03 2 2 0.41 99.26 0.29 0.03 Formulation 4 1 2 0.40 99.26 0.330.01 2 2 0.35 99.42 0.21 0.02

For samples stored at 4° C. for two months, there is virtually no lossin monomer seen by SEC at either site (Tables 23). The amounts ofoligomer are nearly identical for all formulations.

When the different stress conditions are examined (agitation, F/T, andphotostability testing), little degradation is seen by SEC. Agitationseems to cause no appreciable chemical degradation and slight increasesin oligomer levels, as does repeated F/T cycling. All formulationsperform roughly the same. Upon exposure to light, the SEC analysesindicate higher fragmentation levels for formulation 3 (Table 26).

TABLE 25 Overall stability profile as measured by SEC for MOR6654Bsamples stored at 4° C. for two months. Formulation Time RRT 4 C. RepPoint 0.81 0.84 0.88 0.91 1.00 1.07 1.22 1.38 1.58 Formulation 1 1 00.13 99.74 0.04 0.09 2 0 0.14 99.78 0.03 0.04 Formulation 2 1 0 0.3199.64 0.05 2 0 0.33 99.60 0.07 Formulation 3 1 0 0.17 99.78 0.05 0.05 20 0.19 99.77 0.05 Formulation 4 1 0 0.12 99.71 0.17 2 0 0.17 99.59 0.070.17 Formulation 1 1 2 0.32 99.62 0.06 2 2 0.32 99.63 0.05 Formulation 21 2 0.34 99.61 0.05 2 2 0.35 99.59 0.06 Formulation 3 1 2 0.35 99.580.07 2 2 0.36 99.57 0.07 Formulation 4 1 2 0.34 99.60 0.05 2 2 0.3799.57 0.06

TABLE 26 Overall stability profile as measured by SEC for MOR6654Bformulations subjected to agitation stress (agit), multiple F/T cycles(f/t), and prolonged exposure to light (photo). Time RRT Formulation RepPoint 0.81 0.88 0.95 1.00 1.07 1.22 1.29 1.38 1.40 1.58 Formulation 1 10 0.13 99.74 0.04 0.09 2 0 0.14 99.78 0.03 0.04 Formulation 2 1 0 0.3199.64 0.05 2 0 0.33 99.60 0.07 Formulation 3 1 0 0.17 99.78 0.05 0.05 20 0.19 99.77 0.05 Formulation 4 1 0 0.12 99.71 0.17 2 0 0.17 99.59 0.070.17 Formulation 1 1 agit 0.69 99.31 Formulation 2 1 agit 0.94 99.06Formulation 3 1 agit 0.98 98.89 0.11 0.02 Formulation 4 1 agit 0.8599.15 Formulation 1 1 F/T 0.94 98.85 0.20 2 F/T 0.94 98.91 0.15Formulation 2 1 F/T 0.90 98.96 0.13 2 F/T 0.90 99.00 0.09 Formulation 31 F/T 0.92 98.99 0.09 2 F/T 0.92 98.99 0.09 Formulation 4 1 F/T 0.7999.09 0.12 2 F/T 0.79 99.09 0.10 Formulation 1 1 photo 0.77 98.94 0.250.04 2 photo 0.80 98.92 0.25 0.04 Formulation 2 1 photo 0.87 98.86 0.240.03 2 photo 0.90 98.84 0.24 0.03 Formulation 3 1 photo 0.61 0.26 98.440.09 0.57 0.03 2 photo 0.62 0.25 98.49 0.10 0.52 0.02 Formulation 4 1photo 0.91 98.68 0.36 0.05 2 photo 0.95 98.56 0.44 0.05

RP HPLC

RP HPLC provides information on chemical degradation that might beoccurring in the protein.

As chemical degradation seems to be occurring in MOR6654B, the RP HPLCanalysis could be quite informative.

At t₀, all four formulations have a purity near 99.9% by RP HPLC.

After one month storage at 40° C., the purity decreases to approx. 96%,although the purity was markedly higher for formulation 4. The disparityin these values calls into questions whether this value is correct(Table 27).

At two months, all four formulations have decreased to about 94%, withlittle, if any, differences between the formulations.

TABLE 27 Purity of MOR6654B formulations determined by RP HPLC afterstorage at 40° C. for up to two months Form [protein] No mg/mL t0 t1 t21 150 sucrose 99.86 99.89 96.09 93.97 94.01 2 150 trehalose 99.94 99.9296.27 94.61 94.12 3 150 sucrose/Arg 99.93 99.93 95.82 94.13 94.30 4 20sucrose 99.85 99.84 98.44 94.44 94.67

TABLE 28 Purity of MOR6654B formulations determined by RP HPLC afterstorage at 4° C. or 25° C. for two months Form [protein] No mg/mL t0 t24° C. t2 25 C. 1 150 sucrose 99.86 99.89 96.92 96.85 96.63 96.61 2 150trehalose 99.94 99.92 97.12 96.99 96.95 96.63 3 150 sucrose/Arg 99.9399.93 96.88 96.99 96.49 96.46 4 20 sucrose 99.85 99.84 96.81 97.07 96.3496.41

Storage at 25° C. for two months produces a decrease in purity by RPHPLC to ˜96.5% (Table 28). The purity for samples stored at 4° C. wasnot that different, with purities near 97%. Again, all four formulationsperformed similarly.

Upon being subjected to stress conditions, the purity also decreases.For the agitation and F/T studies, the purity is about 97-98% for all ofthe formulations. However, upon exposure to light there is greaterextent of degradation. The 150 mg/mL formulations decrease to about 94%while the 20 mg/mL formulation decreases all the way to <90%.

TABLE 29 Purity determined by RP HPLC of MOR6654B formulations subjectedto agitation stress (agit), multiple F/T cycles (F/T), and prolongedexposure to light (photo). Form [protein] No mg/mL t0 agit F/T photo 1150 sucrose 99.86 99.89 97.51 98.14 94.47 2 150 trehalose 99.94 99.9297.46 97.67 94.45 3 150 sucrose/Arg 99.93 99.93 97.89 96.68 94.82 4 20sucrose 99.85 99.84 96.74 97.29 89.51

As with SEC, it is helpful to examine the appearance of degradationproducts as well as consider the loss of the main peak. For samplesstored at 40° C., the degradation profile is summarized in Table 30.Only one impurity is seen at t0, but multiple species are observed uponstorage at elevated temperature. It appears that the relatively highpurity seen for formulation 4 at t1 was simply due to not seeing thepeak at RRT 1.19 (Table 30). Overall, there is very little differencebetween any of the four formulations regarding overall stabilityprofiled as measured by RP HPLC.

TABLE 30 Degradation profile of MOR6654B formulations determined by RPHPLC after storage at 40° C. for one month (bold) and two months(italic) Formulation Time 40 C. Rep Point 0.67 0.70 0.76 0.86 0.93 1.001.11 1.16 1.19 Formulation 1 1 0 0.14 99.86 2 0 0.11 99.89 Formulation 21 0 0.11 99.89 2 0 0.21 99.79 Formulation 3 1 0 0.12 99.88 2 0 0.1199.89 Formulation 4 1 0 0.15 99.85 2 0 0.16 99.84 Formulation 1 1 1 0.010.04 1.35 96.09 2.51 Formulation 2 1 1 0.01 0.02 1.44 96.33 2.20Formulation 3 1 1 0.02 0.06 1.70 95.99 2.24 Formulation 4 1 1 0.01 0.031.52 98.44 Formulation 1 1 2 0.04 2.72 93.97 1.97 1.29 2 2 0.05 2.7394.01 3.21 Formulation 2 1 2 0.02 2.67 94.61 2.70 2 2 0.06 2.73 94.121.86 1.24 Formulation 3 1 2 0.08 2.87 94.13 2.19 0.73 2 2 0.08 2.8894.30 2.74 Formulation 4 1 2 0.05 2.89 94.44 2.62 2 2 0.05 2.72 94.672.57

For samples stored at 25° C., there are two primary degradation productsthat elute after the main peak (Table 31). The stability profile for allfour formulations is essentially the same at 25° C. when assayed usingRP HPLC. Likewise, there a similar degradation profile for samplesstored at 4° C. (Table 32).

TABLE 31 Degradation profile of MOR6654B formulations determined by RPHPLC after storage at 25° C. for t0 and two months Formulation Time 25C. Rep Point 0.67 0.76 0.86 0.93 1.00 1.11 1.16 1.19 Formulation 1 1 00.14 99.86 2 0 0.11 99.89 Formulation 2 1 0 0.11 99.89 2 0 0.21 99.79Formulation 3 1 0 0.12 99.88 2 0 0.11 99.89 Formulation 4 1 0 0.15 99.852 0 0.16 99.84 Formulation 1 1 2 0.64 96.63 2.13 0.60 2 2 0.56 96.612.24 0.59 Formulation 2 1 2 0.55 96.95 1.77 0.72 2 2 0.56 96.63 2.210.60 Formulation 3 1 2 0.63 96.49 2.33 0.55 2 2 0.65 96.46 2.17 0.72Formulation 4 1 2 0.55 96.34 2.37 0.74 2 2 0.54 96.41 2.16 0.88

TABLE 32 Degradation profile of MOR6654B formulations determined by RPHPLC after storage at 4° C. for up to two months Formulation 4 C. RepTime Point 0.67 0.76 0.86 0.93 1.00 1.11 1.16 1.19 Formulation 1 1 00.14 99.86 2 0 0.11 99.89 Formulation 2 1 0 0.11 99.89 2 0 0.21 99.79Formulation 3 1 0 0.12 99.88 2 0 0.11 99.89 Formulation 4 1 0 0.15 99.852 0 0.16 99.84 Formulation 1 1 2 0.25 96.92 2.20 0.63 2 2 0.24 96.852.34 0.58 Formulation 2 2 2 0.17 97.12 1.87 0.83 2 2 0.23 96.99 1.950.83 Formulation 3 1 2 0.16 96.88 2.96 2 2 0.17 96.99 2.84 Formulation 41 2 0.23 96.81 2.20 0.76 2 2 0.16 97.07 2.06 0.71

When the samples are subjected to agitation, only one degradationproduct appears and the levels are similar in all of the samples (Table33). Formulation 4 seems slightly less stable than the other three,possibly due to the lower protein concentration. Repeated F/T cyclingalso causes some modest damage as seen by RP HPLC, with the profilesimilar to that seen for agitation. This is seen repeatedly throughoutthe study, indicating that the interfacial sensitivity of this proteincan be seen whether one does one test or the other. There does notappear to be a need to conduct both to assess the overall sensitivity tointerfacial stress. Finally, exposure to light crates some early elutingspecies, which are likely fragments. Of the four formulations, thelevels of these species are higher for formulation 3 (Table 33).

TABLE 33 Degradation profile determined by RP HPLC of MOR6654Bformulations subjected to agitation stress (agit), multiple F/T cycles(F/T), and prolonged exposure to light (photo). Formulation AgitationRep Condition 0.68 0.75 0.87 0.93 0.96 1.00 1.10 1.16 1.24 1.29Formulation 1 1 t0 0.14 99.86 2 0.11 99.89 Formulation 2 1 t0 0.11 99.892 0.21 99.79 Formulation 3 1 t0 0.12 99.88 2 0.11 99.89 Formulation 4 1t0 0.15 99.85 2 0.16 99.84 Formulation 1 1 Agit 0.10 98.07 1.83 2 0.1296.96 2.92 Formulation 2 1 Agit 0.29 97.52 2.20 2 0.13 97.40 2.47Formulation 3 1 Agit 0.10 98.14 1.76 2 0.11 97.64 2.25 Formulation 4 1Agit 0.11 96.79 3.10 2 0.11 96.68 3.21 Formulation 1 1 F/T 0.17 99.310.51 2 0.23 96.96 2.81 Formulation 2 1 F/T 0.26 97.94 1.80 2 0.27 97.402.33 Formulation 3 1 F/T 0.17 96.68 3.15 Formulation 4 1 F/T 0.12 97.112.78 2 0.10 97.48 2.42 Formulation 1 1 photo 0.01 0.62 1.17 94.47 3.73 20.01 0.63 1.42 96.93 1.02 Formulation 2 2 photo 0.01 0.65 1.52 94.453.38 2 0.01 0.68 1.56 94.08 3.68 Formulation 3 1 photo 0.03 1.36 0.6294.82 3.17 2 0.03 1.40 0.87 93.98 3.73 Formulation 4 1 photo 0.24 0.986.05 89.51 3.23 2 0.25 0.93 5.45 89.69 3.68

Cationic Exchange Chromatography (CEX HPLC)

The third HPLC method used to evaluate the stability of the fourMOR6654B formulations is cation exchange (CEX) chromatography. Thismethod is intended to identify degradation products that differ incharge from the parent compound. Initially, all four formulationsdisplay a broad main peak that comprises about 98% of the total peakarea (Table 34). Upon storage at 40° C. for one month, the maindecreases to about 96%, with formulation 4 having a slightly higherpurity (Table 34). After two months at 40° C., this has furtherdecreased to about 95%, with formulation 1 showing the highest purity.

TABLE 34 Purity as determined by CEX HPLC of the MOR6654B formulationsafter storage at 40° C. for up to two months Form [protein] t0 t1 t2 Nomg/mL CEX CEX CEX 1 150 sucrose 98.30 98.28 96.76 95.76 95.69 2 150trehalose 98.35 98.28 96.62 95.17 95.31 3 150 sucrose/Arg 98.34 98.2696.62 94.75 94.89 4 20 sucrose 98.30 98.22 97.31 95.39 95.13

TABLE 35 Purity as determined by CEX HPLC of the MOR6654B formulationsafter storage at 4° C. and 25° C. for two months Form [protein] t0 t2 4C. t2 25 C. No mg/mL CEX CEX CEX 1 150 sucrose 98.30 98.28 97.29 97.7097.09 97.36 2 150 trehalose 98.35 98.28 97.45 97.35 97.31 97.20 3 150sucrose/Arg 98.34 98.26 97.33 97.45 97.36 97.35 4 20 sucrose 98.30 98.2297.05 97.38 97.42 97.46

For MOR6654B formulations stored for two months at 25° C., the puritydecreases from >98% to ˜97%, with formulation 4 being slightly, but onlyslightly, more stable (Table 35). When stored at 4° C. for the sameamount of time, there is some loss in purity, similar to what was seenwith the RP HPLC data. Again, it is no clear why there is roughly asmuch degradation in the 4° C. samples as in the 25° C., but the extentof degradation is small and all of the MOR6654B formulations behaveapproximately, the same.

TABLE 36 Purity as determined by CEX HPLC of the MOR6654B formulationssubjected to agitation stress (agit), multiple F/T cycles (F/T), andprolonged exposure to light (photo). Form [protein] t0 agit F/T photo Nomg/mL CEX CEX CEX CEX 1 150 sucrose 98.30 98.28 97.59 97.54 98.91 2 150trehalose 98.35 98.28 97.37 97.49 98.82 3 150 sucrose/Arg 98.34 98.2697.33 97.96 98.41 4 20 sucrose 98.30 98.22 97.31 97.56 98.82

Upon agitation of repeated F/T cycling, there is some loss based on CEXHPLC, with all four formulations performing equally (Table 36).Prolonged exposure to light produced very little damage by CEX HPLC, incontrast to what has been seen with SEC and RP HPLC (Tables 26 and 29,respectively). This suggests that the degradation products generated bylight do not differ in terms of charge from the parent compound.

Reducing capillary Electrophoresis (rCE-SDS)

The capillary electrophoresis method here allows one to look at theextent of damage to the light chain (LC) and heavy chain (HC) of theantibody independently. In addition, it can provide an estimate of howmuch of the protein is not intact LC and HC, as indicated by thenon-LC/HC content. At t0, there is approximately 25% LC and 70% HC bypeak area, not correcting for the differences in size (Table 37), withthe remainder (˜5%) counted as non-LC/HC species (Table 38).

TABLE 37 Percentage of LC and HC as determined by rCE-SDS in MOR6654Bformulations stored at 40°C. for up to two months Form [protein] t0 t1t2 No mg/mL LC HC LC HC LC HC LC HC LC HC 1 150 sucrose 25.0 69.8 25.569.4 26.3 64.4 27.5 62.7 27.6 62.3 2 150 trehalose 25.9 69.4 26.0 69.226.9 64.7 27.0 62.3 27.0 61.3 3 150 sucrose/Arg 24.1 70.2 24.5 69.6 27.662.8 27.5 61.7 27.2 63.8 4 20 sucrose 24.9 67.8 24.9 67.9 27.3 67.5 26.565.0 27.0 63.6

After storage for one month at 40 C, the relative amounts of LC and HChave changed, with LC now accounting for ˜27% and HC between 62 and 68%(Table 37). This indicates that HC is being lost. This is reflected in amarked increase in the amount of non-LC/HC species (Table 38), wherethese amounts have nearly doubled from pre-storage levels.

TABLE 38 Percentage of non-LC/HC species as determined by rCE-SDS inMOR6654B formulations stored at 40° C. for up to two months t0 t1 t2Form non- non- non- No [protein] LC/HC LC/HC LC/HC 1 150 sucrose 5.2 9.39.9 2 150 trehalose 4.8 8.4 11.2 3 150 sucrose/Arg 5.8 9.6 9.9 4 20sucrose 7.3 5.2 9.0

By the end of two months, there is about 10% (or more) of the non-LC/HCspecies. These data suggest that formulation 4 is the most robust, withthe smallest percent increase in non-LC/HC levels.

TABLE 39 Percentage of LC and HC as determined by rCE-SDS in MOR6654Bformulations stored at 4° C. for two months Form [protein] t0 t2 4 C.non- No mg/mL LC HC LC HC LC HC LC HC LC/HC 1 150 sucrose 25.0 69.8 25.569.4 26.4 66.2 26.7 66.8 7.0 2 150 trehalose 25.9 69.4 26.0 69.2 27.368.7 27.3 67.9 4.4 3 150 sucrose/Arg 24.1 70.2 24.5 69.6 27.7 67.3 26.966.3 5.9 4 20 sucrose 24.9 67.8 24.9 67.9 25.8 67.6 25.9 67.3 6.7

When samples are stored at 4° C., there is still some degradation seenby rCE-SDS, as the LC levels rise form ˜25% to ˜27% (Table 39). In thiscase, formulation 2 is the most stable with little increase in theamounts of the non-LC/HC species.

TABLE 40 Percentage of LC and HC as determined by rCE-SDS in MOR6654Bformulations stored at 25° C. for two months Form [protein] t0 t2 25 C.non- No mg/mL LC HC LC HC LC HC LC HC LC/HC 1 150 sucrose 25.0 69.8 25.569.4 27.9 67.4 28.0 67.5 4.6 2 150 trehalose 25.9 69.4 26.0 69.2 27.966.1 27.8 67.5 5.3 3 150 sucrose/Arg 24.1 70.2 24.5 69.6 26.4 67.5 26.966.0 6.6 4 20 sucrose 24.9 67.8 24.9 67.9 26.1 68.0 26.7 68.0 5.6

When stored at 25° C. for two months, there is a comparable increase inLC and decrease in HC content (Table 40). The levels of non-LC/HCspecies do rise, but nearly as much as for the 40 C samples. Of these,formulation 3 appears to have the poorest stability, but the differencesare small.

TABLE 41 Percentage of LC and HC as determined by rCE-SDS in MOR6654Bformulations subjected to agitation stress (agit), multiple F/T cycles(F/T), and prolonged exposure to light (photo). Form [protein] agit non-F/T non- photo non- No mg/mL LC HC LC/HC LC HC LC/HC LC HC LC/HC 1 150sucrose 28.3 67.2 4.5 28.7 67.6 3.7 27.3 66.2 6.5 2 150 trehalose 28.268.3 3.5 27.9 67.7 4.4 27.0 66.1 6.9 3 150 sucrose/Arg 28.3 68.4 3.327.6 69.3 3.1 26.6 66.1 7.3 4 20 sucrose 26.9 69.1 4.0 26.9 69.5 3.626.2 64.6 9.2

The final set of samples to be analyzed by rCE-SDS was those subjectedto the three different stress conditions (Table 41). When samples areagitated or exposed to F/T cycling, there is no appreciable increase innon-LC/HC species for any for the four MOR6654B formulations. This isconsistent with the other findings, that MOR6654B in these formulationsis not very sensitive to interfacial stress. It also reinforces the ideathat these two tests provide comparable assessments of interfacialstability. Prolonged exposure to light did result in some loss of HC andrise in the non-LC/HC levels (Table 41). Of these formulations,formulation 4 showed the biggest changes.

Microflow Imaging (MFI)

Over the last few years there has been an increased focus on the levelsof sub-visible particles in injectable protein products. As a result, anumber of different analytical methods have been developed. One of themost widely known is microflow imaging (MFI). This technique was used tomeasure the sub-visible particle content (in terms of particles per mL)within different size ranges. While data was collected up to 100 um interms of size, only data up to 25 um is presented. The reproducibilityof the MFI data is very good. The average relative standard deviationfor duplicate measurements is about 5%.

TABLE 42 Sub-visible content (in particles/mL) as measured using MFI forMOR6654B formulations stored at 40° C. for one month (in bold) and twomonths (in italics). Samples at t1 were not assayed in duplicate.Otherwise, the values reported are averages ± one standard deviation.Form [protein] No mg/mL Excipient total 1-2 um 2-5 um 5-10 um 10-25 um 1150 sucrose 24137 ± 486 18257 ± 391 4968 ± 33 518 ± 6   78 ± 16 2 150trehalose 12614 ± 660  9580 ± 661 2298 ± 64 568 ± 47 148 ± 10 3 150sucrose/Arg 33374 ± 975 23593 ± 718  8554 ± 174 1134 ± 88  82 ± 5 4 20sucrose 4130 ± 28  2463 ± 134 1197 ± 15 441 ± 76  26 ± 14 1 150 sucrose46508 36658 7768 1684 326 2 150 trehalose 22154 14920 5153 1867 214 3150 sucrose/Arg 34800 22990 9791 1926  82 4 20 sucrose 27372 15663 96601943  98 1 150 sucrose  35130 ± 2897  21931 ± 7725  8703 ± 544 3793 ±596 683 ± 34 2 150 trehalose  24843 ± 3887  19692 ± 3789 4246 ± 39 757 ±27 107 ± 1  3 150 sucrose/Arg  46220 ± 3752 33038 ± 849  10853 ± 19572000 ± 110  239 ± 109 4 20 sucrose  51835 ± 1703 33847 ± 905 14942 ± 74 2411 ± 6  127 ± 11

Often, data summarized for MFI report only the total particle count (inparticles per mL), but this can be misleading, as the numbers aredominated by particles being counted in the 1-2 um size range. In thissize range, silicone oil and air bubbles are prominent, and skew thecounting away form protein-based particles. While these numbers areprovided, it is best to look at particles larger than 2 um, possiblygreater than 5 um in size. Also, for samples reporting averages andstandard deviations, these are the results of duplicate runs.

When stored at 40° C., the sub-visible particle count does rise for allof the formulations (Table 42). Of particular note is formulation 4,where the total particle count is less than 5000 particles per mL at t0,much lower than for the other higher concentration, formulations. Afterone month, all of the formulations now exceed 20000 particles per mL,although the increase for formulation 3 is quite small (Table 42). Atthe two months, only formulation 2 is less than 30000 particles per mL.The difference is even more striking for particles from 2-5 um (wherethis formulation contains less than 5000 particles per mL) and 5-10 um,where the levels are less than 1000 particles per mL, with onlyformulation 1 being close in terms of the total amount of sub-visibleparticles present.

TABLE 43 Sub-visible content (in particles/mL) as measured using MFI forMOR6654B formulations stored at 25° C. for two months (in bold). Thevalues reported are averages ± one standard deviation. Form [protein] Nomg/mL Excipient total 1-2 um 2-5 um 5-10 um 10-25 um 1 150 sucrose 24137± 486 18257 ± 391  4968 ± 33  518 ± 6   78 ± 16 2 150 trehalose 12614 ±660 9580 ± 661 2298 ± 64  568 ± 47 148 ± 10 3 150 sucrose/Arg 33374 ±975 23593 ± 718  8554 ± 174 1134 ± 88  82 ± 5 4 20 sucrose 4130 ± 282463 ± 134 1197 ± 15  441 ± 76  26 ± 14 1 150 sucrose 21362 ± 600 14928± 192  5420 ± 648  907 ± 124 101 ± 20 2 150 trehalose  35948 ± 190024189 ± 2258 9257 ± 252 2416 ± 609  70 ± 13 3 150 sucrose/Arg  67603 ±1743 49362 ± 2758 12381 ± 154  4303 ± 282 1243 ± 560 4 20 sucrose  48163± 2420 31446 ± 950  15394 ± 1356 1240 ± 126  83 ± 13

When MOR6654B samples are stored at 25° C. for two months, there is anincrease in sub-visible particles in all formulations, although theincrease in formulation 1 is very small (Table 43). For formulation 1,there is only a modest increase in particles from 5 to 10 um in size.Otherwise, there is virtually no change. Meanwhile, there is a sizableincrease in all of the other formulations, especially in the 2 to 25 umsize ranges. In particular, formulations 3 and 4 show significantincreases in most size range bins.

For MOR6654B formulations stored at 4° C. for two months there is littlechange in the sub-visible particle levels. Formulation 1 shows only asmall increase in particles from 5-10 mm, with slight decrease in all ofthe other size ranges. Formulation 2 shows only small increases acrossthe different size ranges, while formulation 3 displays small decreasesin the overall particle levels. By comparison, formulation 4, whichstarted with a very low sub-visible particle burden (barely above thatof pure water), shows a significant increase in the particle per mL inall size ranges.

When MOR6654B formulations are subjected to the stress conditions ofagitation, F/T cycling and light exposure, there are large increasesonly for the photostability samples (Table 44). Both agitation and F/Tcycling cause only small to modest increases in particle levels,especially if one ignores particles below 2 um. On the other hand,prolonged light exposure does cause sub-visible particle formation toincreases sizably (Table 44). This is particularly true for formulations3 and 4. By comparison, formulation 1 shows almost no change in theoverall sub-visible particle burden.

TABLE 44 Sub-visible content (in particles/mL) as measured using MFI forMOR6654B formulations subjected to agitation stress (in bold), multipleF/T cycles (in italics), and prolonged exposure to light (underlined).The reported values for t0 are averages ± one standard deviation. Form[protein] No mg/mL Excipients total 1-2 um 2-5 um 5-10 um 10-25 um 1 150Sucrose/PS20/His 24137 ± 486 18257 ± 391 4968 ± 33 518 ± 6  78 ± 16 2150 trehalose/PS20/His 12614 ± 660  9580 ± 661 2298 ± 64 568 ± 47 148 ±10  3 150 sucrose/Arg/PS20/His 33374 ± 975 23593 ± 718  8554 ± 174 1134± 88  82 ± 5  4  20 sucrose/PS20/His 4130 ± 28  2463 ± 134 1197 ± 15 441± 76 26 ± 14 1 150 sucrose/PS20/His 22306 14913 6288 1071  33 2 150trehalose/PS20/His 35220 21637 9932 3345 301 3 150 sucrose/Arg/PS20/His26857 16889 7264 2428 220 4  20 sucrose/PS20/His 10528  6511 3355  576 85 1 150 sucrose/PS20/His 35027 26159 7278 1343 226 2 150trehalose/PS20/His 23502 17580 4856  979  89 3 150 sucrose/Arg/PS20/His21686 15257 5256 1104  66 4  20 sucrose/PS20/His  9667  6078 2198 1340 45 1 150 sucrose/PS20/His 25417 18711 5723  864 102 2 150trehalose/PS20/His 29238 22107 6160  823 147 3 150 sucrose/Ara/PS20/186398  167017  16314  2790 270 His 4  20 sucrose/PS20/His 46159 2935914428  1224 131 His (histidine); P20 (polysorbate 20); Arg (arginine)

Colorimetry

The last analytical method that was used in this stability study wascolorimetry. At t0, the four formulations all display a brown color(Table 45), with a slightly different color intensity for the moredilute formulation (formulation 4). Incubation at 40° C. for one or twomonths leads to little change in color, although formulation 4 does showsome small changes (Table 45). When stored at lower temperatures, thereis little, if any, change in color (Table 46). When subjected to any ofthe stress conditions, the color stays the same as well (Table 47).

TABLE 45 Color of the MOR6654B formulations stored at 40° C. for up totwo months [pro- Form tein] No mg/mL t0 t1 t2 1 150 sucrose B6 B7 B6 B6B6 B6 2 150 trehalose B7 B7 B7 B6 B6 B6 3 150 sucrose/ B6 B6 B6 B6 B6 B6Arg 4 20 sucrose B8 B8 BY7 BY7 B7 B6

TABLE 46 Color of the MOR6654B formulations stored at 4° C. and 25° C.for zero and two months [pro- Form tein] No mg/ml t0 t2 4° C. t2 25° C.1 150 sucrose B6 B7 B7 B7 B6 B6 2 150 trehalose B7 B7 B7 B7 B7 B7 3 150sucrose/ B6 B6 B6 B6 B6 B6 Arg 4 20 sucrose B8 B8 B8 B9 B8 B8

TABLE 47 Color of the MOR6654B formulations subjected to agitationstress (agit), multiple F/T cycles (F/T), and prolonged exposure tolight (photo). [pro- Form tein] No mg/ml t0 agit F/T photo 1 150 sucroseB6 B7 B7 B7 B6 B7 B6 B6 2 150 trehalose B7 B7 B7 B7 B7 B7 B6 B6 3 150sucrose/ B6 B6 B6 B6 B6 B6 B6 B6 Arg 4 20 sucrose B8 B8 B8 B8 B8 B8 B7B7

SUMMARY

A wide variety of biophysical and biochemical analytical methods wasused to determine if there were differences between the formulations interms of stability. Among the formulations selected for the secondscreen, little difference was seen using many of the analyticaltechniques. Only storage at 40° C. and prolonged exposure to lightcaused any significant amount of degradation. While all fourformulations were very similar in terms of their stability profile,formulation 1 (150 mg/mL MOR6654B, 220 mM sucrose, 20 mM histidine,0.04% polysorbate 20, pH 6.0) showed the least propensity to degradeoverall as determined by this battery of analytical methods. Formulation1 was more stable in thermal and light stress conditions, less prone todegradation and was also more stable in terms of particles formation inthe sub-visible range.

1. A method for delivering an anti-BAFFR antibody to a mammal,comprising administering to said mammal an aqueous composition having apH of 5.0-7.0 and comprising (i) an anti-BAFFR antibody wherein theantibody has a concentration of 18-165 mg/mL, and wherein saidanti-BAFFR antibody includes heavy chain CDR1, CDR2 and CDR3 of SEQ IDNOs 3, 4 and 5 respectively, and light chain CDR1, CDR2 and CDR3 of SEQID NOs: 6, 7 and 8, (ii) a stabilizer, (iii) a buffering agent, (iv) asurfactant, and, optionally, (v) an amino acid.
 2. The method of claim1, wherein the patient has a disease or disorder mediated by BAFFreceptor.
 3. The method of claim 1, wherein the patient has a B cellneoplasm.
 4. The method of claim 3, wherein the B cell neoplasm islymphoma, leukemia or myeloma.
 5. A method of killing or depleting Bcells in a mammal, comprising administering to said mammal an aqueouscomposition having a pH of 5.0-7.0 and comprising (i) an anti-BAFFRantibody wherein the antibody has a concentration of 18-165 mg/mL, andwherein said anti-BAFFR antibody includes heavy chain CDR1, CDR2 andCDR3 of SEQ ID NOs 3, 4 and 5 respectively, and light chain CDR1, CDR2and CDR3 of SEQ ID NOs: 6, 7 and 8, (ii) a stabilizer, (iii) a bufferingagent, (iv) a surfactant, and, optionally, (v) an amino acid.
 6. Themethod of claim 5, wherein the mammal is human.